Production of carotenoids in oleaginous yeast and fungi

ABSTRACT

The present invention provides systems for producing engineered oleaginous yeast or fungi that express retinolic compounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of application Ser. No. 12/443,362, filed Mar. 27, 2009, which is a national phase application under 35 U.S.C. §371 of International Application No. PCT/US2007/021092, filed Sep. 28, 2007, which claims the benefit of U.S. provisional patent application No. 60/848,062, filed Sep. 28, 2006, and which claims the benefit of U.S. provisional patent application No. 60/902,145, filed Feb. 16, 2007. The entire contents of each of these applications are hereby incorporated by reference.

SEQUENCE LISTING

In accordance with 37 CFR 1.52(e)(5), a Sequence Listing in the form of a text file (entitled “2005862-0073_SL.txt,” created on Feb. 18, 2014, and is 329,074 bytes in size) is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

Carotenoids are organic pigments ranging in color from yellow to red that are naturally produced by certain organisms, including photosynthetic organisms (e.g., plants, algae, cyanobacteria), and some fungi. Carotenoids are responsible for the orange color of carrots, as well as the pink in flamingos and salmon, and the red in lobsters and shrimp. Animals, however, cannot produce carotenoids and must receive them through their diet.

Carotenoid pigments (e.g., β-carotene and astaxanthin) are used industrially as ingredients for food and feed stocks, both serving a nutritional function and enhancing consumer acceptability. For example, astaxanthin is widely used in salmon aquaculture to provide the orange coloration characteristic of their wild counterparts. Some carotenoids are also precursors of vitamin A. Also, carotenoids have antioxidant properties, and may have various health benefits (see, for example, Jyonouchi et al., Nutr. Cancer 16:93, 1991; Giovannucci et al., J. Natl. Cancer Inst. 87:1767, 1995; Miki, Pure Appl. Chem 63:141, 1991; Chew et al., Anticancer Res. 19:1849, 1999; Wang et al., Antimicrob. Agents Chemother. 44:2452, 2000). Some carotenoids such as β-carotene, lycopene, and lutein are currently sold as nutritional supplements.

In general, the biological systems that produce carotenoids are industrially intractable and/or produce the compounds at such low levels that commercial scale isolation is not practicable. Thus, most carotenoids used in industry are produced by chemical synthesis. There is a need for improved biological systems that produce carotenoids. Some efforts have previously been made to genetically engineer certain bacteria or fungi to produce higher levels of carotenoids (see, for example, Misawa et al., J. Biotechnol. 59:169, 1998; Visser et al., FEMS Yeast Research 4:221, 2003). However, improved systems, allowing higher levels of production and greater ease of isolation, are needed.

SUMMARY OF THE INVENTION

The present invention provides improved systems for the biological production of carotenoids and/or retinolic compounds. In one aspect, the invention encompasses the discovery that it is desirable to produce carotenoids and/or retinolic compounds in oleaginous organisms. Without wishing to be bound by any particular theory, the present inventors propose that biological systems may be able to accumulate higher levels of carotenoids and/or retinolic compounds if the compounds are sequestered in lipid bodies. Regardless of whether absolute levels are higher, however, carotenoids and/or retinolic compounds that are accumulated within lipid bodies in oleaginous organisms are readily isolatable through isolation of the lipid bodies.

The present invention therefore provides oleaginous fungi (including, for example, yeast) that produce one or more carotenoids and/or retinolic compounds. The present invention also provides methods of constructing such yeast and fungi, methods of using such yeast and fungi to produce carotenoids and/or retinolic compounds, and methods of preparing carotenoid-containing compositions and/or retinolic compound-containing compositions, such as food or feed additives, or nutritional supplements, using carotenoids and/or retinolic compounds produced in such oleaginous yeast or fungi. In particular, the present invention provides systems and methods for generating yeast and fungi containing one or more oleaginic and/or carotenogenic and/or retinologenic modifications that increase the oleaginicity and/or alter their carotenoid-producing and/or retinolic compound-producing capabilities as compared with otherwise identical organisms that lack the modification(s).

The present invention further encompasses the general recognition that lipid-accumulating systems are useful for the production and/or isolation of lipophilic agents (such as, but not limited to isoprenoids, or isoprenoid-derived compounds such as retinolic compounds, carotenoids, ubiquinones, lanosterol, zymosterol, ergosterol, vitamins (e.g., vitamins A, E, D, K, specifically 7-dehydrocholesterol (provitamin D3), sterols (e.g., squalene), etc.). According to the present invention, it is desirable to engineer organisms to produce such lipophilic agents and/or to accumulate lipid.

Indeed, one aspect of the present invention is the recognition that host cells can be engineered to accumulate in lipid bodies any of a variety of hydrophilic and/or fat soluble compounds (e.g., retinolic compounds, carotenoids, ubiquinones, vitamins, squalene, etc.) having negligible solubility in water (whether hot or cold) and an appropriate solubility in oil. In some embodiments of the invention, modified host cells are engineered to produce one or more lipophilic agents characterized by negligible solubility in water and detectable solubility in one or more oils. In some embodiments, such lipophilic agents (including, but not limited to carotenoids and/or retinolic compounds) have a solubility in oil below about 0.2%. In some embodiments, such lipophilic agents have a solubility in oil within the range of about <0.001%-0.2%.

The present invention therefore provides engineered host cells (and methods of making and using them) that contain lipid bodies and that further contain one or more compounds accumulated in the lipid bodies, where the compounds are characterized by a negligible solubility in water and a solubility in oil within the range of about <0.001%-0.2%; 0.004%-0.15%; 0.005-0.1%; or 0.005-0.5%. For example, in some embodiments, such lipophilic agents have a solubility in oil below about 0.15%, 0.14%, 0.13%, 0.12%, 0.11%, 0.10%. 0.09, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.05%, or less. In some embodiments, the lipophilic agents show such solubility in an oil selected from the group consisting of sesame; soybean; apricot kernel; palm; peanut; safflower; coconut; olive; cocoa butter; palm kernel; shea butter; sunflower; almond; avocado; borage; carnauba; hazel nut; castor; cotton seed; evening primrose; orange roughy; rapeseed; rice bran; walnut; wheat germ; peach kernel; babassu; mango seed; black current seed; jojoba; macademia nut; sea buckthorn; sasquana; tsubaki; mallow; meadowfoam seed; coffee; emu; mink; grape seed; thistle; tea tree; pumpkin seed; kukui nut; and mixtures thereof.

In some embodiments, the present invention provides a recombinant fungus. In certain embodiments, the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight; and produces at least one carotenoid and/or retinolic compound, and can accumulate the produced carotenoid and/or retinolic compound to at least about 1% of its dry cell weight; wherein the recombinant fungus comprises at least one modification as compared with a parental fungus, which parental fungus both is not oleaginous and does not accumulate the carotenoid and/or retinolic compound to at least about 1% of its dry cell weight, the at least one modification being selected from the group consisting of retinologenic modifications, carotenogenic modifications, oleaginic modifications, and combinations thereof, and wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one carotenoid and/or retinolic compound to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one carotenoid and/or retinolic compound which the parental fungus does not produce.

In other embodiments, the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight; and produces at least one carotenoid selected from the group consisting of antheraxanthin, adonirubin, adonixanthin, astaxanthin, canthaxanthin, capsorubrin, β-cryptoxanthin, α-carotene, β,ψ-carotene, δ-carotene, ε-carotene, echinenone, 3-hydroxyechinenone, 3′-hydroxyechinenone, γ-carotene, ψ-carotene, 4-keto-γ-carotene, ζ-carotene, α-cryptoxanthin, deoxyflexixanthin, diatoxanthin, 7,8-didehydroastaxanthin, didehydrolycopene, fucoxanthin, fucoxanthinol, isorenieratene, β-isorenieratene, lactucaxanthin, lutein, lycopene, myxobactone, neoxanthin, neurosporene, hydroxyneurosporene, peridinin, phytoene, rhodopin, rhodopin glucoside, 4-keto-rubixanthin, siphonaxanthin, spheroidene, spheroidenone, spirilloxanthin, torulene, 4-keto-torulene, 3-hydroxy-4-keto-torulene, uriolide, uriolide acetate, violaxanthin, zeaxanthin-β-diglucoside, zeaxanthin, a C30 carotenoid, and combinations thereof, and can accumulate the produced carotenoid to at least about 1% of its dry cell weight; wherein the recombinant fungus comprises at least one modification as compared with a parental fungus, the at least one modification being selected from the group consisting of carotenogenic modifications, oleaginic modifications, and combinations thereof, and wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one carotenoid to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one carotenoid which the parental fungus does not naturally produce.

In other embodiments, the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight; and produces at least one retinolic compound selected from the group consisting of retinol, retinal, retinoic acid, and combinations thereof, and can accumulate the produced retinolic compound to at least about 1% of its dry cell weight; wherein the recombinant fungus comprises at least one modification as compared with a parental fungus, the at least one modification being selected from the group consisting of retinologenic modifications, carotenogenic modifications, oleaginic modifications, and combinations thereof, and wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one retinolic compound to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one retinolic compound which the parental fungus does not naturally produce.

In some embodiments, the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight; and produces at least one carotenoid and/or retinolic compound, and can accumulate the produced carotenoid and/or retinolic compound to at least about 1% of its dry cell weight; wherein the recombinant fungus is a member of a genus selected from the group consisting of: Aspergillus, Blakeslea, Botrytis, Candida, Cercospora, Cryptococcus, Cunninghamella, Fusarium (Gibberella), Kluyveromyces, Lipomyces, Mortierella, Mucor, Neurospora, Penicillium, Phycomyces, Pichia (Hansenula), Puccinia, Pythium, Rhodosporidium, Rhodotorula, Saccharomyces, Sclerotium, Trichoderma, Trichosporon, Xanthophyllomyces (Phaffia), and Yarrowia; or is a species selected from the group consisting of: Aspergillus terreus, Aspergillus nidulans, Aspergillus niger, Blakeslea trispora, Botrytis cinerea, Candida japonica, Candida pulcherrima, Candida revkaufi, Candida tropicalis, Candida utilis, Cercospora nicotianae, Cryptococcus curvatus, Cunninghamella echinulata, Cunninghamella elegans, Fusarium fujikuroi (Gibberella zeae), Kluyveromyces lactis, Lipomyces starkeyi, Lipomyces lipoferus, Mortierella alpina, Mortierella ramanniana, Mortierella isabellina, Mortierella vinacea, Mucor circinelloides, Neurospora crassa, Phycomyces blakesleanus, Pichia pastoris, Puccinia distincta, Pythium irregulare, Rhodosporidium toruloides, Rhodotorula glutinis, Rhodotorula graminis, Rhodotorula mucilaginosa, Rhodotorula pinicola, Rhodotorula gracilis, Saccharomyces cerevisiae, Sclerotium rolfsii, Trichoderma reesei, Trichosporon cutaneum, Trichosporon pullans, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), and Yarrowia lipolytica, wherein the recombinant fungus comprises at least one modification as compared with a parental fungus, the at least one modification being selected from the group consisting of retinologenic modifications, carotenogenic modifications, oleaginic modifications, and combinations thereof, and wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one carotenoid and/or retinolic compound to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one carotenoid and/or retinolic compound which the parental fungus does not naturally produce.

In other embodiments, the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight; and produces at least one carotenoid selected from the group consisting of antheraxanthin, adonirubin, adonixanthin, astaxanthin, canthaxanthin, capsorubrin, β-cryptoxanthin, α-carotene, β,ψ-carotene, δ-carotene, ε-carotene, echinenone, 3-hydroxyechinenone, 3′-hydroxyechinenone, γ-carotene, ψ-carotene, 4-keto-γ-carotene, ζ-carotene, α-cryptoxanthin, deoxyflexixanthin, diatoxanthin, 7,8-didehydroastaxanthin, didehydrolycopene, fucoxanthin, fucoxanthinol, isorenieratene, β-isorenieratene, lactucaxanthin, lutein, lycopene, myxobactone, neoxanthin, neurosporene, hydroxyneurosporene, peridinin, phytoene, rhodopin, rhodopin glucoside, 4-keto-rubixanthin, siphonaxanthin, spheroidene, spheroidenone, spirilloxanthin, torulene, 4-keto-torulene, 3-hydroxy-4-keto-torulene, uriolide, uriolide acetate, violaxanthin, zeaxanthin-β-diglucoside, zeaxanthin, a C30 carotenoid, and combinations thereof, and can accumulate the produced carotenoid to at least about 1% of its dry cell weight; wherein the recombinant fungus is a member of a genus selected from the group consisting of: Aspergillus, Blakeslea, Botrytis, Candida, Cercospora, Cryptococcus, Cunninghamella, Fusarium (Gibberella), Kluyveromyces, Lipomyces, Mortierella, Mucor, Neurospora, Penicillium, Phycomyces, Pichia (Hansenula), Puccinia, Pythium, Rhodosporidium, Rhodotorula, Saccharomyces, Sclerotium, Trichoderma, Trichosporon, Xanthophyllomyces (Phaffia), and Yarrowia, or is of a species selected from the group consisting of Aspergillus terreus, Aspergillus nidulans, Aspergillus niger, Blakeslea trispora, Botrytis cinerea, Candida japonica, Candida pulcherrima, Candida revkaufi, Candida tropicalis, Candida utilis, Cercospora nicotianae, Cryptococcus curvatus, Cunninghamella echinulata, Cunninghamella elegans, Fusarium fujikuroi (Gibberella zeae), Kluyveromyces lactis, Lipomyces starkeyi, Lipomyces lipoferus, Mortierella alpina, Mortierella ramanniana, Mortierella isabellina, Mortierella vinacea, Mucor circinelloides, Neurospora crassa, Phycomyces blakesleanus, Pichia pastoris, Puccinia distincta, Pythium irregulare, Rhodosporidium toruloides, Rhodotorula glutinis, Rhodotorula graminis, Rhodotorula mucilaginosa, Rhodotorula pinicola, Rhodotorula gracilis, Saccharomyces cerevisiae, Sclerotium rolfsii, Trichoderma reesei, Trichosporon cutaneum, Trichosporon pullans, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), and Yarrowia lipolytica, wherein the recombinant fungus comprises at least one modification as compared with a parental fungus, the at least one modification being selected from the group consisting of carotenogenic modifications, oleaginic modifications, and combinations thereof, and wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one carotenoid to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one carotenoid which the parental fungus does not naturally produce.

In other embodiments, the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight; and produces at least one retinolic compound selected from the group consisting of retinol, retinal, retinoic acid, and combinations thereof, and can accumulate the produced retinolic compound to at least about 1% of its dry cell weight; wherein the recombinant fungus is a member of a genus selected from the group consisting of: Aspergillus, Blakeslea, Botrytis, Candida, Cercospora, Cryptococcus, Cunninghamella, Fusarium (Gibberella), Kluyveromyces, Lipomyces, Mortierella, Mucor, Neurospora, Penicillium, Phycomyces, Pichia (Hansenula), Puccinia, Pythium, Rhodosporidium, Rhodotorula, Saccharomyces, Sclerotium, Trichoderma, Trichosporon, Xanthophyllomyces (Phaffia), and Yarrowia, or is of a species selected from the group consisting of Aspergillus terreus, Aspergillus nidulans, Aspergillus niger, Blakeslea trispora, Botrytis cinerea, Candida japonica, Candida pulcherrima, Candida revkaufi, Candida tropicalis, Candida utilis, Cercospora nicotianae, Cryptococcus curvatus, Cunninghamella echinulata, Cunninghamella elegans, Fusarium fujikuroi (Gibberella zeae), Kluyveromyces lactis, Lipomyces starkeyi, Lipomyces lipoferus, Mortierella alpina, Mortierella ramanniana, Mortierella isabellina, Mortierella vinacea, Mucor circinelloides, Neurospora crassa, Phycomyces blakesleanus, Pichia pastoris, Puccinia distincta, Pythium irregulare, Rhodosporidium toruloides, Rhodotorula glutinis, Rhodotorula graminis, Rhodotorula mucilaginosa, Rhodotorula pinicola, Rhodotorula gracilis, Saccharomyces cerevisiae, Sclerotium rolfsii, Trichoderma reesei, Trichosporon cutaneum, Trichosporon pullans, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), and Yarrowia lipolytica, wherein the recombinant fungus comprises at least one modification as compared with a parental fungus, the at least one modification being selected from the group consisting of retinologenic modifications, carotenogenic modifications, oleaginic modifications, and combinations thereof, and wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one retinolic compound to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one retinolic compound which the parental fungus does not naturally produce.

In certain embodiments, the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight; and the recombinant fungus produces at least one small molecule lipophilic agent selected from the group consisting of retinolic compounds, carotenoids, ubiquinone, vitamin K, vitamin E, squalene, lanosterol, zymosterol, ergosterol, 7-dehydrocholesterol (provitamin D3), and combinations thereof and can accumulate the produced carotenoid and/or retinolic compound to at least about 1% of its dry cell weight; wherein the recombinant fungus comprises at least one modification as compared with a parental fungus, the at least one modification being selected from the group consisting of retinologenic modifications, carotenogenic modifications, oleaginic modifications, and combinations thereof, and wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one carotenoid and/or retinolic compound to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one carotenoid and/or retinolic compound which the parental fungus does not naturally produce.

In some embodiments, the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight; and the recombinant fungus produces at least one small molecule lipophilic agent characterized by a negligible solubility in water and solubility in oil within the range of about <0.001%-0.2%; 0.004%-0.15%; 0.005-0.1%; or 0.005-0.5%, and combinations thereof and can accumulate the produced small molecule lipophilic agent to at least about 1% of its dry cell weight; wherein the recombinant fungus comprises at least one modification as compared with a parental fungus, the at least one modification being selected from the group consisting of retinologenic modifications, carotenogenic modifications, oleaginic modifications, and combinations thereof, and wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one small molecule lipophilic agent to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one small molecule lipophilic agent which the parental fungus does not naturally produce.

In other embodiments the recombinant fungus is oleaginous in that it can accumulate lipid to at least about 20% of its dry cell weight; and the recombinant fungus produces at least one small molecule lipophilic agent selected from the group consisting of retinolic compounds, carotenoids, ubiquinone, vitamin K, vitamin E, squalene, lanosterol, zymosterol, ergosterol, 7-dehydrocholesterol (provitamin D3), and can accumulate the produced small molecule lipophilic agent to at least about 1% of its dry cell weight; wherein the recombinant fungus is a member of a genus selected from the group consisting of Candida, Cryptococcus, Cunninghamella, Lipomyces, Mortierella, Mucor, Phycomyces, Pythium, Rhodosporidium, Rhodotorula, Trichosporon, Yarrowia, Aspergillus, Botrytis, Cercospora, Fusarium (Gibberella), Kluyveromyces, Neurospora, Penicillium, Pichia (Hansenula), Puccinia, Saccharomyces, Schizosaccharomyces, Sclerotium, Trichoderms, Ustilago, and Xanthophyllomyces (Phaffia) and comprises at least one genetic modification as compared with a parental fungus, wherein the at least one modification alters oleaginicity of the recombinant fungus, confers to the recombinant fungus oleaginy, confers to the recombinant fungus the ability to produce the at least one small molecule lipophilic agent to a level at least about 1% of its dry cell weight, or confers to the recombinant fungus the ability to produce at least one small molecule lipophilic agent which the parental fungus does not naturally produce.

In some embodiments, the present invention provides a strain of Yarrowia lipolytica comprising one or more modifications selected from the group consisting of an oleaginic modification, a carotenogenic modification, and combinations thereof, such that the strain accumulates from 1% to 15% of its dry cell weight as at least one carotenoid. In some embodiments, the present invention provides a strain of Yarrowia lipolytica comprising one or more modifications selected from the group consisting of an oleaginic modification, a retinologenic modification, and combinations thereof, such that the strain accumulates from 1% to 15% of its dry cell weight as at least one retinolic compound.

In some embodiments, the present invention provides an engineered Y. lipolytica strain that produces β-carotene, the strain containing one or more carotenogenic modifications selected from the group consisting of: increased expression or activity of a Y. lipolytica GGPP synthase polypeptide; expression or activity of a truncated HMG CoA reductase polypeptide; expression or activity of a phytoene dehydrogenase polypeptide; expression or activity of a phytoene synthase/lycopene cyclase polypeptide; increased expression or activity of an FPP synthase polypeptide; increased expression or activity of an IPP isomerase polypeptide; increased expression or activity of an HMG synthase polypeptide; increased expression or activity of a mevalonate kinase polypeptide; increased expression or activity of a phosphomevalonate kinase polypeptide; increased expression or activity of a mevalonate pyrophosphate decarboxylate polypeptide; increased expression or activity of a malic enzyme polypeptide; increased expression or activity of a malate dehydrogenase polypeptide; increased expression or activity of an AMP deaminase polypeptide; increased expression or activity of a glucose 6 phosphate dehydrogenase polypeptide; increased expression or activity of a malate dehydrogenase homolog2 polypeptide; increased expression or activity of a GND1-6-phosphogluconate dehydrogenase polypeptide; increased expression or activity of a isocitrate dehydrogenase polypeptide; increased expression or activity of a IDH2-isocitrate dehydrogenase polypeptide; increased expression or activity of a fructose 1,6 bisphosphatase polypeptide; increased expression or activity of a Erg10-acetoacetyl CoA thiolase polypeptide; increased expression or activity of a ATP citrate lyase subunit 2 polypeptide; increased expression or activity of a ATP citrate lyase subunit 1 polypeptide; decreased expression or activity of a squalene synthase polypeptide; decreased expression or activity of a prenyldiphosphate synthase polypeptide; or decreased expression or activity of a PHB polyprenyltransferase polypeptide; and combinations thereof.

In some embodiments, the present invention provides an engineered Y. lipolytica strain that produces Vitamin A, the strain containing one or more retinologenic modifications selected from the group consisting of: increased expression or activity of a beta-carotene 15,15′-monooxygenase polypeptide; increased expression or activity of a retinol dehydrogenase polypeptide; and combinations thereof.

In some embodiments, the present invention provides an engineered Y. lipolytica strain containing a truncated HMG CoA reductase polypeptide. In some embodiments, the present invention provides an engineered Y. lipolytica strain having increased expression or activity of a GGPP synthase gene. In some embodiments, the present invention provides an engineered Y. lipolytica strain having decreased expression or activity of a squalene synthase polypeptide. In some embodiments, the present invention provides an engineered Y. lipolytica strain containing a heterologous phytoene dehydrogenase (carB) polypeptide and a heterologous phytoene synthase/lycopene cyclase (carRP) polypeptide.

In some embodiments, the present invention provides a genetically modified Y. lipolytica strain comprising an altered activity or expression of one or more enzymes when compared to an unmodified strain, wherein the altered activity or expression of one or more enzymes is selected from the group consisting of: increased activity or expression of a beta-carotene 15,15′-monooxygenase polypeptide; increased activity or expression of a retinol dehydrogenase polypeptide; increased activity or expression of acetyl-CoA thiolase, increased activity or expression of HMG-CoA synthase, increased activity or expression of HMG-CoA reductase, increased activity or expression of mevalonate kinase, increased activity or expression of phosphomevalonate kinase, increased activity or expression of mevalonate PP decarboxylase, decreased activity or expression of acetyl-CoA carboxylase, increased activity or expression of IPP isomerase, increased activity or expression of GPP synthase, increased activity or expression of FPP synthase, increased activity or expression of squalene synthase, decreased activity or expression of squalene synthase, increased activity or expression of GGPP synthase, decreased activity or expression of GGPP synthase, increased activity or expression of glucose-6-phosphate dehydrogenase, increased activity or expression of 6-phosphogluconate dehydrogenase, increased activity or expression of fructose 1,6 bisphosphatase, increased activity or expression of NADH kinase, increased activity or expression of transhydrogenase, and combinations thereof.

In certain embodiments, the present invention provides a genetically modified Candida utilis strain comprising an altered activity or expression of one or more enzymes when compared to an unmodified strain, wherein the altered activity or expression of one or more enzymes is selected from the group consisting of: increased activity or expression of a beta-carotene 15,15′-monooxygenase polypeptide; increased activity or expression of a retinol dehydrogenase polypeptide; increased activity or expression of acetyl-CoA thiolase, increased activity or expression of HMG-CoA synthase, increased activity or expression of HMG-CoA reductase, increased activity or expression of mevalonate kinase, increased activity or expression of phosphomevalonate kinase, increased activity or expression of mevalonate PP decarboxylase, decreased activity or expression of acetyl-CoA carboxylase, increased activity or expression of IPP isomerase, increased activity or expression of GPP synthase, increased activity or expression of FPP synthase, increased activity or expression of squalene synthase, decreased activity or expression of squalene synthase, increased activity or expression of GGPP synthase, decreased activity or expression of GGPP synthase, increased activity or expression of glucose-6-phosphate dehydrogenase, increased activity or expression of 6-phosphogluconate dehydrogenase, increased activity or expression of fructose 1,6 bisphosphatase, increased activity or expression of NADH kinase, increased activity or expression of transhydrogenase, and combinations thereof.

In other embodiments, the present invention provides a genetically modified Saccharomyces cerevisiae strain comprising an altered activity or expression of one or more enzymes when compared to an unmodified strain, wherein the altered activity or expression of one or more enzymes is selected from the group consisting of: increased activity or expression of a beta-carotene 15,15′-monooxygenase polypeptide; increased activity or expression of a retinol dehydrogenase polypeptide; increased activity or expression of acetyl-CoA thiolase, increased activity or expression of HMG-CoA synthase, increased activity or expression of HMG-CoA reductase, increased activity or expression of mevalonate kinase, increased activity or expression of phosphomevalonate kinase, increased activity or expression of mevalonate PP decarboxylase, decreased activity or expression of acetyl-CoA carboxylase, increased activity or expression of IPP isomerase, increased activity or expression of GPP synthase, increased activity or expression of FPP synthase, increased activity or expression of squalene synthase, decreased activity or expression of squalene synthase, increased activity or expression of GGPP synthase, decreased activity or expression of GGPP synthase, increased activity or expression of glucose-6-phosphate dehydrogenase, increased activity or expression of 6-phosphogluconate dehydrogenase, increased activity or expression of fructose 1,6 bisphosphatase, increased activity or expression of NADH kinase, increased activity or expression of transhydrogenase, and combinations thereof.

In some embodiments, the present invention provides a genetically modified Xanthophyllomyces dendrorhous (Phaffia rhodozyma) strain comprising an altered activity or expression of one or more enzymes when compared to an unmodified strain, wherein the altered activity or expression of one or more enzymes is selected from the group consisting of: increased activity or expression of a beta-carotene 15,15′-monooxygenase polypeptide; increased activity or expression of a retinol dehydrogenase polypeptide; increased activity or expression of acetyl-CoA thiolase, increased activity or expression of HMG-CoA synthase, increased activity or expression of HMG-CoA reductase, increased activity or expression of mevalonate kinase, increased activity or expression of phosphomevalonate kinase, increased activity or expression of mevalonate PP decarboxylase, decreased activity or expression of acetyl-CoA carboxylase, increased activity or expression of IPP isomerase, increased activity or expression of GPP synthase, increased activity or expression of FPP synthase, increased activity or expression of squalene synthase, decreased activity or expression of squalene synthase, increased activity or expression of GGPP synthase, decreased activity or expression of GGPP synthase, increased activity or expression of glucose-6-phosphate dehydrogenase, increased activity or expression of 6-phosphogluconate dehydrogenase, increased activity or expression of fructose 1,6 bisphosphatase, increased activity or expression of NADH kinase, increased activity or expression of transhydrogenase, and combinations thereof.

In other embodiments, the present invention provides a method of producing a carotenoid, the method comprising steps of cultivating a fungus under conditions that allow production of the carotenoid; and isolating the produced carotenoid. In other embodiments, the present invention provides a method of producing a retinolic compound, the method comprising steps of cultivating a fungus under conditions that allow production of the retinolic compound; and isolating the produced retinolic compound.

In certain embodiments, the present invention provides an isolated carotenoid composition, prepared by a method comprising steps of cultivating the fungus under conditions that allow production of a carotenoid; and isolating the produced carotenoid. In certain embodiments, the present invention provides an isolated retinolic compound composition, prepared by a method comprising steps of cultivating the fungus under conditions that allow production of a retinolic compound; and isolating the produced retinolic compound.

In other embodiments, the present invention provides a composition comprising lipid bodies; at least one carotenoid compound; and intact fungal cells. In other embodiments, the present invention provides a composition comprising lipid bodies; at least one retinolic compound; and intact fungal cells.

In some embodiments, the present invention provides a composition comprising: an oil suspension comprising: lipid bodies; at least one carotenoid compound; intact fungal cells; and a binder or filler. In some embodiments, the present invention provides a composition comprising: an oil suspension comprising: lipid bodies; at least one retinolic compound; intact fungal cells; and a binder or filler.

In certain embodiments, the present invention provides a composition comprising: an oil suspension comprising: lipid bodies; at least one carotenoid compound; intact fungal cells; and one or more other agents selected from the group consisting of chelating agents, pigments, salts, surfactants, moisturizers, viscosity modifiers, thickeners, emollients, fragrances, preservatives, and combinations thereof. In certain embodiments, the present invention provides a composition comprising: an oil suspension comprising: lipid bodies; at least one retinolic compound; intact fungal cells; and one or more other agents selected from the group consisting of chelating agents, pigments, salts, surfactants, moisturizers, viscosity modifiers, thickeners, emollients, fragrances, preservatives, and combinations thereof.

In some embodiments, the present invention provides a feedstuff comprising a carotenoid in lipid bodies. In other embodiments, the present invention provides a feedstuff comprising a carotenoid in lipid bodies; wherein the carotenoid is selected from the group consisting of astaxanthin, β-carotene, canthaxanthin, zeaxanthin, lutein, lycopene, echinenone, β-cryptoxanthin and combinations thereof. In some embodiments, the present invention provides a feedstuff comprising a retinolic compound in lipid bodies. In other embodiments, the present invention provides a feedstuff comprising a retinolic compound in lipid bodies; wherein the retinolic compound is selected from the group consisting of retinol, retainal, retinoic acid, and combinations thereof.

In certain embodiments, the present invention provides a carotenoid composition comprising a Y. lipolytica cell containing at least 1% carotenoids by weight. In other embodiments, the present invention provides a carotenoid composition comprising Y. lipolytica lipid bodies; and at least one carotenoid compound, wherein the at least one carotenoid compound is present at a level that is at least 1% by weight of the lipid bodies. In certain embodiments, the present invention provides a retinolic compound composition comprising a Y. lipolytica cell containing at least 1% retinolic compounds by weight. In other embodiments, the present invention provides a retinolic compound composition comprising Y. lipolytica lipid bodies; and at least one retinolic compound, wherein the at least one retinolic compound is present at a level that is at least 1% by weight of the lipid bodies.

Additional aspects of the present invention will be apparent to those of ordinary skill in the art from the present description, including the appended claims.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1A-1D depicts certain common carotenoids.

FIG. 2 depicts how sufficient levels of acetyl-CoA and NADPH may be accumulated in the cytosol of oleaginous organisms to allow for production of significant levels of cytosolic lipids. Enzymes: 1, pyruvate decarboxylase; 2, malate dehydrogenase; 3, malic enzyme; 4, pyruvate dehydrogenase; 5, citrate synthase; 6, ATP-citrate lyase; 7, citrate/malate translocase.

FIGS. 3A and 3B depict the mevalonate isoprenoid biosynthesis pathway, which typically operates in eukaryotes, including fungi.

FIG. 4 depicts the mevalonate-independent isoprenoid biosynthesis pathway, also known as the DXP pathway, which typically operates in bacteria and in the plastids of plants.

FIG. 5 depicts intermediates in the isoprenoid biosynthesis pathway and how they feed into biosynthetic pathways of other biomolecules, including carotenoids as well as non-carotenoid compounds such as sterols, steroids, and vitamins, such as vitamin E or vitamin K.

FIGS. 6A-6D illustrate various carotenoid biosynthetic pathways. FIG. 6A highlights branches leading to various cyclic and acyclic xanthophylls; FIG. 6B shows certain X dendrorhous pathways that generate dicyclic and monocyclic carotenoids, including astaxanthin; FIG. 6C shows interconnecting pathways for converting β-carotene into any of a variety of other carotenoids, including astaxanthin; FIG. 6D depicts possible routes of synthesis of cyclic carotenoids and common plant and algal xanthophylls from neurosporene.

FIG. 7 and subparts FIGS. 7A-7I show an alignment of certain representative fungal HMG-CoA reductase polypeptides (SEQ ID NOs 176-182, respectively, in order of appearance). As can be seen, these polypeptides show very high identity across the catalytic region, and also have complex membrane spanning domains. In some embodiments of the invention, these membrane-spanning domains are disrupted or are removed, so that, for example, a hyperactive version of the polypeptide may be produced.

FIGS. 8A-8P depict schematic representations of plasmids generated and described in detail in the exemplification.

FIGS. 9A-E show production characteristics of certain engineered cells according to the present invention. Specifically, Panel A shows β-Carotene and phytoene production by Strain MF760 when grown in glycerol, glucose or olive oil; Panel B shows dry cell weight accumulation of strain MF760 when grown in glycerol, glucose or olive oil; Panel C shows β-Carotene and dry cell weight analysis of strain MF760 when grown in the presence of a combination of olive oil and glucose; Panel D shows canthaxanthin, echinenone and β-carotene production of strain MF840; and Panel E shows canthaxanthin and echinenone production of strain MF838 in a 2 phase feeding protocol.

FIG. 10 is a Table listing certain Y. lipolytica genes representing various polypeptides (e.g., oleaginic and isoprenoid biosynthesis peptides) useful in engineering cells in accordance with the present invention. FIG. 10 discloses SEQ ID NOs 183-218, respectively, in order of appearance.

FIG. 11 depicts the all-trans-retinol (Vitamin A) biosynthesis pathway, starting with beta-carotene as a substrate.

FIGS. 12a-e depict various characteristics of strain ML1011 (MF740 transformed with multiple integrated copies of the X. autotrophicus crtZ gene) grown under different pH conditions. FIG. 12a depicts accumulation of total carotenoid (absorbance units per unit dry cell weight) over the course of the fermentation. FIG. 12b depicts accumulation of zeaxanthin (absorbance units per dry cell weight; AU) over the course of the fermentation. FIG. 12c depicts the fraction of carotenoid as zeaxanthin (AU zeaxanthin/AU total carotenoid) over the course of the fermentation. FIG. 12d depicts carbon dioxide evolution over the course of the fermentation. FIG. 12e depicts biomass accumulation over the course of the fermentation.

DEFINITIONS

Aromatic amino acid biosynthesis polypeptide: The term “aromatic amino acid biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of aromatic amino acids in yeast and/or bacteria through chorismate and the shikimate pathway. For example, as discussed herein, anthranilate synthase, enzymes of the shikimate pathway, chorismate mutase, chorismate synthase, DAHP synthase, and transketolase are all aromatic amino acid biosynthesis polypeptides. Each of these polypeptides is also a ubiquinone biosynthesis polypeptide or a ubiquinone biosynthesis competitor for purposes of the present invention, as production of chorismate is a precursor in the synthesis of para-hydroxybenzoate for the biosynthesis of ubiquinone.

Biosynthesis polypeptide: The term “biosynthesis polypeptide” as used herein (typically in reference to a particular compound or class of compounds), refers to polypeptides involved in the production of the compound or class of compounds. In some embodiments of the invention, biosynthesis polypeptides are synthetic enzymes that catalyze particular steps in a synthesis pathway that ultimately produces a relevant compound. In some embodiments, the term “biosynthesis polypeptide” may also encompass polypeptides that do not themselves catalyze synthetic reactions, but that regulate expression and/or activity of other polypeptides that do so. Biosynthesis polypeptides include, for example, aromatic amino acid biosynthesis polypeptides, C₅₋₉ quinone biosynthesis polypeptides, carotenoid biosynthesis polypeptides, retinolic compound biosynthesis polypeptides, FPP biosynthesis polypeptides, isoprenoid biosynthesis polypeptides, PHB biosynthesis polypeptides, quinone biosynthesis polypeptides, sterol biosynthesis polypeptides, ubiquinone biosynthesis polypeptides, Vitamin D biosynthesis polypeptides, Vitamin E biosynthesis polypeptides, and Vitamin K biosynthesis polypeptides.

C₅₋₉ quinone biosynthesis polypeptide: The term “C₅₋₉ quinone biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of a C₅₋₉ quinone, for example a polyprenyldiphosphate synthase polypeptide. To mention but a few, these include, for example, pentaprenyl, hexaprenyl, heptaprenyl, octaprenyl, and/or solanesyl (nonaprenyl) diphosphate synthase polypeptides (i.e., polypeptides that perform the chemical reactions performed by the pentaprenyl, hexaprenyl, heptaprenyl, octaprenyl, and solanesyl (nonaprenyl) polypeptides, respectively (see also Okada et al., Biochim. Biophys. Acta 1302:217, 1996; Okada et al., J. Bacteriol. 179:5992, 1997). As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, C₅₋₉ quinone biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other C₅₋₉ quinone biosynthesis polypeptides.

Carotenogenic modification: The term “carotenogenic modification”, as used herein, refers to a modification of a host organism that adjusts production of one or more carotenoids, as described herein. For example, a carotenogenic modification may increase the production level of one or more carotenoids, and/or may alter relative production levels of different carotenoids. In principle, an inventive carotenogenic modification may be any chemical, physiological, genetic, or other modification that appropriately alters production of one or more carotenoids in a host organism produced by that organism as compared with the level produced in an otherwise identical organism not subject to the same modification. In most embodiments, however, the carotenogenic modification will comprise a genetic modification, typically resulting in increased production of one or more selected carotenoids. In some embodiments, the carotenogenic modification comprises at least one chemical, physiological, genetic, or other modification; in other embodiments, the carotenogenic modification comprises more than one chemical, physiological, genetic, or other modification. In certain aspects where more than one modification is utilized, such modifications can comprise any combination of chemical, physiological, genetic, or other modification (e.g., one or more genetic, chemical, and/or physiological modification(s)). In some embodiments, the selected carotenoid is one or more of astaxanthin, β-carotene, canthaxanthin, lutein, lycopene, phytoene, zeaxanthin, and/or modifications of zeaxanthin or astaxanthin (e.g., glucoside, esterified zeaxanthin or astaxanthin). In some embodiments, the selected carotenoid is one or more xanthophylls, and/or a modification thereof (e.g., glucoside, esterified xanthophylls). In certain embodiments, the selected xanthophyll is selected from the group consisting of astaxanthin, lutein, zeaxanthin, lycopene, and modifications thereof. In some embodiments, the selected carotenoid is one or more of astaxanthin, β-carotene, canthaxanthin, lutein, lycopene, and zeaxanthin and/or modifications of zeaxanthin or astaxanthin. In some embodiments, the carotenoid is β-carotene. In some embodiments, the selected carotenoid is astaxanthin. In some embodiments, the selected carotenoid is other than β-carotene.

Carotenogenic polypeptide: The term “carotenogenic polypeptide”, as used herein, refers to any polypeptide that is involved in the process of producing carotenoids in a cell, and may include polypeptides that are involved in processes other than carotenoid production but whose activities affect the extent or level of production of one or more carotenoids, for example by scavenging a substrate or reactant utilized by a carotenoid polypeptide that is directly involved in carotenoid production. Carotenogenic polypeptides include isoprenoid biosynthesis polypeptides, carotenoid biosynthesis polypeptides, and isoprenoid biosynthesis competitor polypeptides, as those terms are defined herein. The term also encompasses polypeptides that may affect the extent to which carotenoids are accumulated in lipid bodies.

Carotenoid: The term “carotenoid” is understood in the art to refer to a structurally diverse class of pigments derived from isoprenoid pathway intermediates. The commitment step in carotenoid biosynthesis is the formation of phytoene from geranylgeranyl pyrophosphate. Carotenoids can be acyclic or cyclic, and may or may not contain oxygen, so that the term carotenoids include both carotenes and xanthophylls. In general, carotenoids are hydrocarbon compounds having a conjugated polyene carbon skeleton formally derived from the five-carbon compound IPP, including triterpenes (C₃₀ diapocarotenoids) and tetraterpenes (C₄₀ carotenoids) as well as their oxygenated derivatives and other compounds that are, for example, C₃₅, C₅₀, C₆₀, C₇₀, C₈₀ in length or other lengths. Many carotenoids have strong light absorbing properties and may range in length in excess of C₂₀₀. C₃₀ diapocarotenoids typically consist of six isoprenoid units joined in such a manner that the arrangement of isoprenoid units is reversed at the center of the molecule so that the two central methyl groups are in a 1,6-positional relationship and the remaining non-terminal methyl groups are in a 1,5-positional relationship. Such C₃₀ carotenoids may be formally derived from the acyclic C₃₀H₄₂ structure, having a long central chain of conjugated double bonds, by: (i) hydrogenation (ii) dehydrogenation, (iii) cyclization, (iv) oxidation, (v) esterification/glycosylation, or any combination of these processes. C₄₀ carotenoids typically consist of eight isoprenoid units joined in such a manner that the arrangement of isoprenoid units is reversed at the center of the molecule so that the two central methyl groups are in a 1,6-positional relationship and the remaining non-terminal methyl groups are in a 1,5-positional relationship. Such C₄₀ carotenoids may be formally derived from the acyclic C₄₀H₅₆ structure, having a long central chain of conjugated double bonds, by (i) hydrogenation, (ii) dehydrogenation, (iii) cyclization, (iv) oxidation, (v) esterification/glycosylation, or any combination of these processes. The class of C₄₀ carotenoids also includes certain compounds that arise from rearrangements of the carbon skeleton, or by the (formal) removal of part of this structure. More than 600 different carotenoids have been identified in nature; certain common carotenoids are depicted in FIG. 1. Carotenoids include but are not limited to: antheraxanthin, adonirubin, adonixanthin, astaxanthin, canthaxanthin, capsorubrin, β-cryptoxanthin, α-carotene, β-carotene, β,ψ-carotene, δ-carotene, ε-carotene, echinenone, 3-hydroxyechinenone, 3′-hydroxyechinenone, γ-carotene, ψ-carotene, 4-keto-γ-carotene, ζ-carotene, α-cryptoxanthin, deoxyflexixanthin, diatoxanthin, 7,8-didehydroastaxanthin, didehydrolycopene, fucoxanthin, fucoxanthinol, isorenieratene, β-isorenieratene, lactucaxanthin, lutein, lycopene, myxobactone, neoxanthin, neurosporene, hydroxyneurosporene, peridinin, phytoene, rhodopin, rhodopin glucoside, 4-keto-rubixanthin, siphonaxanthin, spheroidene, spheroidenone, spirilloxanthin, torulene, 4-keto-torulene, 3-hydroxy-4-keto-torulene, uriolide, uriolide acetate, violaxanthin, zeaxanthin-β-diglucoside, zeaxanthin, and C30 carotenoids. Additionally, carotenoid compounds include derivatives of these molecules, which may include hydroxy-, methoxy-, oxo-, epoxy-, carboxy-, or aldehydic functional groups. Further, included carotenoid compounds include ester (e.g., glycoside ester, fatty acid ester) and sulfate derivatives (e.g., esterified xanthophylls).

Carotenoid biosynthesis polypeptide: The term “carotenoid biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of one or more carotenoids. To mention but a few, these carotenoid biosynthesis polypeptides include, for example, polypeptides of phytoene synthase, phytoene dehydrogenase (or desaturase), lycopene cyclase, carotenoid ketolase, carotenoid hydroxylase, astaxanthin synthase, carotenoid epsilon hydroxylase, lycopene cyclase (beta and epsilon subunits), carotenoid glucosyltransferase, and acyl CoA:diacylglycerol acyltransferase. In some instances, a single gene may encode a protein with multiple carotenoid biosynthesis polypeptide activities. Representative examples of carotenoid biosynthesis polypeptide sequences are presented in Tables 17a-25. As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, carotenoid biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other carotenoid biosynthesis polypeptides.

FPP biosynthesis polypeptides: The term “FPP biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of farnesyl pyrophosphate. As discussed herein, farnesyl pyrophosphate represents the branchpoint between the sterol biosynthesis pathway and the carotenoid and other biosynthesis pathways. One specific example of an FPP biosynthesis polypeptide is FPP synthase. Representative examples of FPP synthase polypeptide sequences are presented in Table 14. As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, FPP biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other FPP biosynthesis polypeptides.

Gene: The term “gene”, as used herein, generally refers to a nucleic acid encoding a polypeptide, optionally including certain regulatory elements that may affect expression of one or more gene products (i.e., RNA or protein).

Heterologous: The term “heterologous”, as used herein to refer to genes or polypeptides, refers to a gene or polypeptide that does not naturally occur in the organism in which it is being expressed. It will be understood that, in general, when a heterologous gene or polypeptide is selected for introduction into and/or expression by a host cell, the particular source organism from which the heterologous gene or polypeptide may be selected is not essential to the practice of the present invention. Relevant considerations may include, for example, how closely related the potential source and host organisms are in evolution, or how related the source organism is with other source organisms from which sequences of other relevant polypeptides have been selected. Where a plurality of different heterologous polypeptides are to be introduced into and/or expressed by a host cell, different polypeptides may be from different source organisms, or from the same source organism. To give but one example, in some cases, individual polypeptides may represent individual subunits of a complex protein activity and/or may be required to work in concert with other polypeptides in order to achieve the goals of the present invention. In some embodiments, it will often be desirable for such polypeptides to be from the same source organism, and/or to be sufficiently related to function appropriately when expressed together in a host cell. In some embodiments, such polypeptides may be from different, even unrelated source organisms. It will further be understood that, where a heterologous polypeptide is to be expressed in a host cell, it will often be desirable to utilize nucleic acid sequences encoding the polypeptide that have been adjusted to accommodate codon preferences of the host cell and/or to link the encoding sequences with regulatory elements active in the host cell. For example, when the host cell is a Yarrowia strain (e.g., Yarrowia lipolytica), it will often be desirable to alter the gene sequence encoding a given polypeptide such that it conforms more closely with the codon preferences of such a Yarrowia strain. In certain embodiments, a gene sequence encoding a given polypeptide is altered to conform more closely with the codon preference of a species related to the host cell. For example, when the host cell is a Yarrowia strain (e.g., Yarrowia lipolytica), it will often be desirable to alter the gene sequence encoding a given polypeptide such that it conforms more closely with the codon preferences of a related fungal strain. Such embodiments are advantageous when the gene sequence encoding a given polypeptide is difficult to optimize to conform to the codon preference of the host cell due to experimental (e.g., cloning) and/or other reasons. In certain embodiments, the gene sequence encoding a given polypeptide is optimized even when such a gene sequence is derived from the host cell itself (and thus is not heterologous). For example, a gene sequence encoding a polypeptide of interest may not be codon optimized for expression in a given host cell even though such a gene sequence is isolated from the host cell strain. In such embodiments, the gene sequence may be further optimized to account for codon preferences of the host cell. Those of ordinary skill in the art will be aware of host cell codon preferences and will be able to employ inventive methods and compositions disclosed herein to optimize expression of a given polypeptide in the host cell.

Host cell: As used herein, the “host cell” is a fungal cell or yeast cell that is manipulated according to the present invention to accumulate lipid and/or to express one or more carotenoids as described herein. A “modified host cell”, as used herein, is any host cell which has been modified, engineered, or manipulated in accordance with the present invention as compared with a parental cell. In some embodiments, the modified host cell has at least one carotenogenic and/or at least one oleaginic modification. In some embodiments, the modified host cell containing at least one oleaginic modification and/or one carotenogenic modification further has at least one sterologenic modification and/or at least one quinonogenic modification. In some embodiments, the parental cell is a naturally occurring parental cell.

Isolated: The term “isolated”, as used herein, means that the isolated entity has been separated from at least one component with which it was previously associated. When most other components have been removed, the isolated entity is “purified” or “concentrated”. Isolation and/or purification and/or concentration may be performed using any techniques known in the art including, for example, fractionation, extraction, precipitation, or other separation.

Isoprenoid biosynthesis competitor: The term “isoprenoid biosynthesis competitor”, as used herein, refers to an agent whose presence or activity in a cell reduces the level of geranylgeranyl diphosphate (GGPP) available to enter the carotenoid biosynthesis pathway. The term “isoprenoid biosynthesis competitor” encompasses both polypeptide and non-polypeptide (e.g., small molecule) inhibitor agents. Those of ordinary skill in the art will appreciate that certain competitor agents that do not act as inhibitors of isoprenoid biosynthesis generally can nonetheless act as inhibitors of biosynthesis of a particular isoprenoid compound. Particular examples of isoprenoid biosynthesis competitor agents act on isoprenoid intermediates prior to GGPP, such that less GGPP is generated (see, for example, FIG. 5). Squalene synthase is but one isoprenoid biosynthesis competitor polypeptide according to the present invention; representative squalene synthase sequences are presented in Table 16. Prenyldiphosphate synthase enzymes and para-hydroxybenzoate (PHB) polyprenyltransferase are yet additional isoprenoid biosynthesis competitor polypeptides according to the present invention; representative prenyldiphosphate synthase enzymes and PHB polyprenyltransferase polypeptides are presented in Tables 29 and 30, respectively. In certain embodiments, one or more polypeptide components of the SAGA complex are isoprenoid biosynthesis competitors according to the present invention. Genes encoding SAGA complex polypeptides are presented in Table 69. In certain embodiments, a polypeptide encoded by these and other SAGA complex genes is an isoprenoid biosynthesis competitor polypeptide according to the present invention. Those of ordinary skill in the art, considering the known metabolic pathways relating to isoprenoid production and/or metabolism (see, for example, FIGS. 3-6 and other Figures and references herein) will readily appreciate a variety of other particular isoprenoid biosynthesis competitors, including isoprenoid biosynthesis polypeptides.

Isoprenoid biosynthesis polypeptide: The term “isoprenoid biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of isoprenoids. For example, as discussed herein, acetoacetyl-CoA thiolase, HMG-CoA synthase, HMG-CoA reductase, mevalonate kinase, phosphomevalonate kinase, mevalonate pyrophosphate decarboxylase, IPP isomerase, FPP synthase, and GGPP synthase, are all involved in the mevalonate pathway for isoprenoid biosynthesis. Each of these proteins is also an isoprenoid biosynthesis polypeptide for purposes of the present invention, and sequences of representative examples of these enzymes are provided in Tables 7-15. As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, isoprenoid biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other isoprenoid biosynthesis polypeptides (e.g., of one or more enzymes that participates in isoprenoid synthesis). Thus, for instance, transcription factors that regulate expression of isoprenoid biosynthesis enzymes can be isoprenoid biosynthesis polypeptides for purposes of the present invention. To give but a couple of examples, the S. cerevisae Upc2 and YLR228c genes, and the Y. lipolytica YALI0B00660g gene encode transcription factors that are isoprenoid biosynthesis polypeptides according to certain embodiments of the present invention. For instance, the semidominant upc2-1 point mutant (G888D) exhibits increases sterol levels (Crowley et al. J Bacteriol. 180: 4177-4183, 1998). Corresponding YLR228c mutants have been made and tested (Shianna et al. J Bacteriology 183:830-834, 2001); such mutants may be useful in accordance with the present invention, as may be YALI0B00660g derivatives with corresponding upc2-1 mutation(s).

Isoprenoid pathway: The term “isoprenoid pathway” is understood in the art to refer to a metabolic pathway that either produces or utilizes the five-carbon metabolite isopentyl pyrophosphate (IPP). As discussed herein, two different pathways can produce the common isoprenoid precursor IPP—the “mevalonate pathway” and the “non-mevalonate pathway”. The term “isoprenoid pathway” is sufficiently general to encompass both of these types of pathway. Biosynthesis of isoprenoids from IPP occurs by polymerization of several five-carbon isoprene subunits. Isoprenoid metabolites derived from IPP are of varying size and chemical structure, including both cyclic and acyclic molecules. Isoprenoid metabolites include, but are not limited to, monoterpenes, sesquiterpenes, diterpenes, sterols, and polyprenols such as carotenoids.

Oleaginic modification: The term “oleaginic modification”, as used herein, refers to a modification of a host organism that adjusts the desirable oleaginy of that host organism, as described herein. In some cases, the host organism will already be oleaginous in that it will have the ability to accumulate lipid to at least about 20% of its dry cell weight. It may nonetheless be desirable to apply an oleaginic modification to such an organism, in accordance with the present invention, for example to increase (or, in some cases, possibly to decrease) its total lipid accumulation, or to adjust the types or amounts of one or more particular lipids it accumulates (e.g., to increase relative accumulation of triacylglycerol). In other cases, the host organism may be non-oleaginous (though may contain some enzymatic and regulatory components used in other organisms to accumulate lipid), and may require oleaginic modification in order to become oleaginous in accordance with the present invention. The present invention also contemplates application of oleaginic modification to non-oleaginous host strains such that their oleaginicity is increased even though, even after being modified, they may not be oleaginous as defined herein. In principle, the oleaginic modification may be any chemical, physiological, genetic, or other modification that appropriately alters oleaginy of a host organism as compared with an otherwise identical organism not subjected to the oleaginic modification. In most embodiments, however, the oleaginic modification will comprise a genetic modification, typically resulting in increased production and/or activity of one or more oleaginic polypeptides. In some embodiments, the oleaginic modification comprises at least one chemical, physiological, genetic, or other modification; in other embodiments, the oleaginic modification comprises more than one chemical, physiological, genetic, or other modification. In certain aspects where more than one modification is utilized, such modifications can comprise any combination of chemical, physiological, genetic, or other modification (e.g., one or more genetic, chemical and/or physiological modification(s)).

Oleaginic polypeptide: The term “oleaginic polypeptide”, as used herein, refers to any polypeptide that is involved in the process of lipid accumulation in a cell and may include polypeptides that are involved in processes other than lipid biosynthesis but whose activities affect the extent or level of accumulation of one or more lipids, for example by scavenging a substrate or reactant utilized by an oleaginic polypeptide that is directly involved in lipid accumulation. For example, as discussed herein, acetyl-CoA carboxylase, pyruvate decarboxylase, isocitrate dehydrogenase, ATP-citrate lyase, malic enzyme, malate dehydrogenase, and AMP deaminase, among other proteins, are all involved in lipid accumulation in cells. In general, reducing the activity of pyruvate decarboxylase or isocitrate dehydrogenase, and/or increasing the activity of acetyl CoA carboxylase, ATP-citrate lyase, malic enzyme, malate dehydrogenase, and/or AMP deaminase is expected to promote oleaginy. Each of these proteins is an oleaginic peptide for the purposes of the present invention, and sequences of representative examples of these enzymes are provided in Tables 1-6, and 30. Other peptides that can be involved in regenerating NADPH may include, for example, 6-phosphogluconate dehydrogenase (gnd); Fructose 1,6 bisphosphatase (fbp); Glucose 6 phosphate dehydrogenase (g6pd); NADH kinase (EC 2.7.1.86); and/or transhydrogenase (EC 1.6.1.1 and 1.6.1.2). Alternative or additional strategies to promote oleaginy may include one or more of the following: (1) increased or heterologous expression of one or more of acyl-CoA:diacylglycerol acyltransferase (e.g., DGA1; YALI0E32769g); phospholipid:diacylglycerol acyltransferase (e.g., LRO1; YALI0E16797g); and acyl-CoA:cholesterol acyltransferase (e.g., ARE genes such as ARE1, ARE2, YALI0F06578g), which are involved in triglyceride synthesis (Kalscheuer et al. Appl Environ Microbiol p. 7119-7125, 2004; Oelkers et al. J Biol Chem 277:8877-8881, 2002; and Sorger et al. J Biol Chem 279:31190-31196, 2004), (2) decreased expression of triglyceride lipases (e.g., TGL3 and/or TGL4; YALI0D17534g and/or YALI0F10010g (Kurat et al. J Biol Chem 281:491-500, 2006); and (3) decreased expression of one or more acyl-coenzyme A oxidase activities, for example encoded by POX genes (e.g. POX1, POX2, POX3, POX4, POX5; YALI0C23859g, YALI0D24750g, YALI0E06567g, YALI0E27654g, YALI0E32835g, YALI0F10857g; see, for example, Mlickova et al. Appl Environ Microbiol 70: 3918-3924, 2004; Binns et al. J Cell Biol 173:719, 2006). Each of these proteins is an oleaginic peptide for the purposes of the present invention, and sequences of representative examples of these enzymes are provided in Tables 31-43 and 45-47.

Oleaginous: The term “oleaginous”, refers to the ability of an organism to accumulate lipid to at least about 20% of its dry cell weight. In certain embodiments of the invention, oleaginous yeast or fungi accumulate lipid to at least about 25% of their dry cell weight. In other embodiments, inventive oleaginous yeast or fungi accumulate lipid within the range of about 20-45% (e.g., about 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, or more) of their dry cell weight. In some embodiments, oleaginous organisms may accumulate lipid to as much as about 70% of their dry cell weight. In some embodiments of the invention, oleaginous organisms may accumulate a large fraction of total lipid accumulation in the form of triacylglycerol. In certain embodiments, the majority of the accumulated lipid is in the form of triacylglycerol. Alternatively or additionally, the lipid may accumulate in the form of intracellular lipid bodies, or oil bodies. In certain embodiments, the present invention utilizes yeast or fungi that are naturally oleaginous. In some aspects, naturally oleaginous organisms are manipulated (e.g., genetically, chemically, or otherwise) so as to further increase the level of accumulated lipid in the organism. In other embodiments, yeast or fungi that are not naturally oleaginous are manipulated (e.g., genetically, chemically, or otherwise) to accumulate lipid as described herein. For example, for the purposes of the present invention, Saccharomyces cerevisiae, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), and Candida utilis are not naturally oleaginous fungi.

PHB polypeptide or PHB biosynthesis polypeptide: The terms “PHB polypeptide” or “PHB biosynthesis polypeptide” as used herein refers to a polypeptide that is involved in the synthesis of para-hydroxybenzoate from chorismate. In prokaryotes and lower eukaryotes, synthesis of para-hydroxybenzoate occurs by the action of chorismate pyruvate lyase. Biosynthesis of para-hydroxybenzoate from tyrosine or phenylalanine occurs through a five-step process in mammalian cells. Lower eukaryotes such as yeast can utilize either method for production of para-hydroxybenzoate. For example, enzymes of the shikimate pathway, chorismate synthase, DAHP synthase, and transketolase are all PHB biosynthesis polypeptides. Each of these polypeptides is also a ubiquinone biosynthesis polypeptide or a ubiquinone biosynthesis competitor polypeptide for purposes of the present invention.

Polypeptide: The term “polypeptide”, as used herein, generally has its art-recognized meaning of a polymer of at least three amino acids. However, the term is also used to refer to specific functional classes of polypeptides, such as, for example, oleaginic polypeptides, carotenogenic polypeptides, isoprenoid biosynthesis polypeptides, carotenoid biosynthesis polypeptides, etc. For each such class, the present specification provides several examples of known sequences of such polypeptides. Those of ordinary skill in the art will appreciate, however, that the term “polypeptide” is intended to be sufficiently general as to encompass not only polypeptides having the complete sequence recited herein (or in a reference or database specifically mentioned herein), but also to encompass polypeptides that represent functional fragments (i.e., fragments retaining at least one activity) of such complete polypeptides. Moreover, those of ordinary skill in the art understand that protein sequences generally tolerate some substitution without destroying activity. Thus, any polypeptide that retains activity and shares at least about 30-40% overall sequence identity, often greater than about 50%, 60%, 70%, or 80%, and further usually including at least one region of much higher identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99% in one or more highly conserved regions (e.g., isocitrate dehydrogenase polypeptides often share a conserved AMP-binding motif; HMG-CoA reductase polypeptides typically include a highly conserved catalytic domain (see, for example, FIG. 7); acetyl coA carboxylase typically has a carboxyl transferase domain; see, for example, Downing et al., Chem. Abs. 93:484, 1980; Gil et al., Cell 41:249, 1985; Jitrapakdee et al. Curr Protein Pept Sci. 4:217, 2003; U.S. Pat. No. 5,349,126, each of which is incorporated herein by reference in its entirety), usually encompassing at least 3-4 and often up to 20 or more amino acids, with another polypeptide of the same class, is encompassed within the relevant term “polypeptide” as used herein. Other regions of similarity and/or identity can be determined by those of ordinary skill in the art by analysis of the sequences of various polypeptides presented in the Tables herein.

Quinone biosynthesis polypeptide: A “quinone biosynthesis polypeptide”, as that term is used herein, refers to any polypeptide involved in the synthesis of one or more quinone derived compound, as described herein. In particular, quinone biosynthesis polypeptides include ubiquinone biosynthesis polypeptides, C₅₋₉ quinone biosynthesis polypeptides, vitamin K biosynthesis polypeptides, and vitamin E biosynthesis polypeptides.

Quinonogenic modification: The term “quinonogenic modification, as used herein, refers to a modification of a host organism that adjusts production of one or more quinone derived compounds (e.g., ubiquinone, vitamin K compounds, vitamin E compounds, etc.), as described herein. For example, a quinonogenic modification may increase the production level of a particular quinone derived compound, or of a variety of different quinone derived compounds. In some embodiments of the invention, production of a particular quinone derived compound may be increased while production of other quinone derived compounds is decreased. In some embodiments of the invention, production of a plurality of different quinone derived compounds is increased. In principle, an inventive quinonogenic modification may be any chemical, physiological, genetic, or other modification that appropriately alters production of one or more quinone derived compounds in a host organism produced by that organism as compared with the level produced in an otherwise identical organism not subject to the same modification. In most embodiments, however, the quinonogenic modification will comprise a genetic modification, typically resulting in increased production of one or more quinone derived compounds (e.g., ubiquinone, vitamin K compounds, vitamin E compounds). In some embodiments, the quinonogenic modification comprises at least one chemical, physiological, genetic, or other modification; in other embodiments, the quinonogenic modification comprises more than one chemical, physiological, genetic, or other modification. In certain aspects where more than one modification is utilized, such modifications can comprise any combination of chemical, physiological, genetic, or other modification (e.g., one or more genetic, chemical and/or physiological modification(s)).

Retinologenic modification: The term “retinologenic modification”, as used herein, refers to a modification of a host organism that adjusts production of one or more retinolic compounds, as described herein. For example, a retinologenic modification may increase the production level of one or more retinolic compounds, and/or may alter relative production levels of different retinolic compounds. In principle, an inventive retinologenic modification may be any chemical, physiological, genetic, or other modification that appropriately alters production of one or more retinolic compounds in a host organism produced by that organism as compared with the level produced in an otherwise identical organism not subject to the same modification. In most embodiments, however, the retinologenic modification will comprise a genetic modification, typically resulting in increased production of one or more selected retinolic compounds. In some embodiments, the retinologenic modification comprises at least one chemical, physiological, genetic, or other modification; in other embodiments, the retinologenic modification comprises more than one chemical, physiological, genetic, or other modification. In certain aspects where more than one modification is utilized, such modifications can comprise any combination of chemical, physiological, genetic, or other modification (e.g., one or more genetic, chemical, and/or physiological modification(s)). In some embodiments, the selected retinolic compound is one or more of retinol, retinal, and retinoic acid. In some embodiments, the selected retinolic compound is retinol or esters of retinol, including but not limited to retinyl palmitate or retinyl acetate. In some embodiments, the selected retinolic compound is retinoic acid. In some embodiments, the selected retinolic compound is other than retinol.

Retinologenic polypeptide: The term “retinologenic polypeptide”, as used herein, refers to any polypeptide that is involved in the process of producing retinolic compounds in a cell, and may include polypeptides that are involved in processes other than retinolic compound production but whose activities affect the extent or level of production of one or more retinolic compounds, for example by scavenging a substrate or reactant utilized by a retinologenic polypeptide that is directly involved in retinolic compound production. Retinologenic polypeptides include retinolic compound biosynthesis polypeptides, isoprenoid biosynthesis polypeptides, carotenoid biosynthesis polypeptides, and isoprenoid biosynthesis competitor polypeptides, as those terms are defined herein. The term also encompasses polypeptides that may affect the extent to which retinolic compounds are accumulated in lipid bodies.

Retinolic compounds: The term “retinolic compound” is understood in the art to refer to a structurally similar class of compounds derived from certain carotenoids, collectively referred to as Vitamin A. All forms of Vitamin A have a beta-ionone ring to which an isoprenoid chain is attached. Retinolic compounds include, for example, retinol (the alcohol form), retinal (the aldehyde form), and retinoic acid (the acid form). Many different geometric isomers of retinol, retinal and retinoic acid are possible as a result of either a trans or cis configuration of four of the five double bonds found in the polyene chain. The cis isomers are less stable and can readily convert to the all-trans configuration. Nevertheless, some cis isomers are found naturally and carry out essential functions. For example, the 11-cis-retinal isomer is the chromophore of rhodopsin, the vertebrate photoreceptor molecule. The term retinolic compound also includes esters of retinol such as retinyl palmitate or retinyl acetate. Hydrolysis of retinyl esters results in retinol. Retinal, also known as retinaldehyde, can be reversibly reduced to produce retinol or it can be irreversibly oxidized to produce retinoic acid. The best described active retinoid metabolites are 11-cis-retinal and the all-trans and 9-cis-isomers of retinoic acid.

Retinolic compound biosynthesis polypeptides: The term “retinolic compound biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of one or more retinolic compounds. To mention but a few, these retinolic compound biosynthesis polypeptides include, for example, polypeptides of beta-carotene 15,15′-monooxygenase (also known as beta-carotene dioxygenase) and/or beta-carotene retinol dehydrogenase. In some instances, a single gene may encode a protein with multiple retinolic compound biosynthesis polypeptide activities. Representative examples of retinolic compound biosynthesis polypeptide sequences are presented in Tables 67 and 68. As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, retinolic compound biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other retinolic compound biosynthesis polypeptides.

Small Molecule: In general, a small molecule is understood in the art to be an organic molecule that is less than about 5 kilodaltons (Kd) in size. In some embodiments, the small molecule is less than about 3 Kd, 2 Kd, or 1 Kd. In some embodiments, the small molecule is less than about 800 daltons (D), 600 D, 500 D, 400 D, 300 D, 200 D, or 100 D. In some embodiments, small molecules are non-polymeric. In some embodiments, small molecules are not proteins, peptides, or amino acids. In some embodiments, small molecules are not nucleic acids or nucleotides. In some embodiments, small molecules are not saccharides or polysaccharides.

Source organism: The term “source organism”, as used herein, refers to the organism in which a particular polypeptide sequence can be found in nature. Thus, for example, if one or more heterologous polypeptides is/are being expressed in a host organism, the organism in which the polypeptides are expressed in nature (and/or from which their genes were originally cloned) is referred to as the “source organism”. Where multiple heterologous polypeptides are being expressed in a host organism, one or more source organism(s) may be utilized for independent selection of each of the heterologous polypeptide(s). It will be appreciated that any and all organisms that naturally contain relevant polypeptide sequences may be used as source organisms in accordance with the present invention. Representative source organisms include, for example, animal, mammalian, insect, plant, fungal, yeast, algal, bacterial, cyanobacterial, archaebacterial and protozoal source organisms.

Sterol biosynthesis polypeptide: The term “sterol biosynthesis polypeptide”, as used herein, refers to any polypeptide that is involved in the synthesis of one or more sterol compounds. Thus, sterol biosynthesis polypeptides can include isoprenoid biosynthesis polypeptides to the extent that they are involved in production of isopentyl pyrophosphate. Moreover, the term refers to any polypeptide that acts downstream of farnesyl pyrophosphate and in involved in the production of one or more sterol compounds. For example, sterol biosynthesis polypeptides include squalene synthase, which catalyses conversion of farnesyl pyrophosphate to presqualene pyrophosphate, and further catalyzes conversion of presqualene pyrophosphate to squalene, e.g., the enzyme with EC number 2.5.1.21. In some embodiments of the invention, sterol biosynthesis polypeptides further include one or more polypeptides involved in metabolizing squalene into a vitamin D compound. Thus, sterol biosynthesis polypeptides can include one or more of the polypeptides designated by EC number 1.14.99.7, 5.4.99.7, 5.4.99.8, 5.3.3.5, 1.14.21.6, 1.14.15.-, and/or 1.14.13.13, as well as other enzyme polypeptides involved in the sterol biosynthesis pathways. Furthermore, sterol biosynthesis polypeptides can include one or more enzyme polypeptides including, for example, C-14 demethylase (ERG9), squalene monooxygenase (ERG1), 2,3-oxidosqualene-lanosterol synthase (ERG7), C-1 demethylase (ERG11), C-14 reductase (ERG24), C-4 methyloxidase (ERG25), C-4 decarboxylase (ERG26), 3-ketoreductase (ERG27), C-24 methyltransferase (ERG6), Δ8-7 isomerase (ERG2), C-5 desaturase (ERG3), C-22 desaturase (ERG5) and/or C-24 reductase (ERG4) polypeptides, and/or other polypeptides involved in producing one or more vitamin D compounds (e.g., vitamin D2, vitamin D3, or a precursor thereof). As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, sterol biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other sterol biosynthesis polypeptides. Thus, for instance, transcription factors that regulate expression of sterol biosynthesis enzymes can be sterol biosynthesis polypeptides for purposes of the present invention. To give but a couple of examples, the S. cerevisiae Upc2 and YLR228c genes, and the Y. lipolytica YALI0B00660g gene encode transcription factors that are sterol biosynthesis polypeptides according to certain embodiments of the present invention. For instance, the semidominant upc2-1 point mutation (G888D) exhibits increased sterol levels (Crowley et al., J. Bacteriol 180:4177-4183, 1998). Corresponding YLR228c mutants have been made and tested (Shianna et al., J Bacteriol 183:830, 2001); such mutants may be useful in accordance with the present invention, as may be YALI0B00660g derivatives with corresponding upc2-1 mutation(s). Representative examples of sterol biosynthesis polypeptide sequences are presented in Tables 53-66. As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, sterol biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other sterol biosynthesis polypeptides.

Sterologenic modification: The term “sterologenic modification”, as used herein, refers to a modification of a host organism that adjusts production of one or more sterol compounds (e.g., squalene, lanosterol, zymosterol, ergosterol, 7-dehydrocholesterol (provitamin D3), vitamin D compound(s), etc.), as described herein. For example, a sterologenic modification may increase the production level of a particular sterol compound, or of a variety of different sterol compounds. In some embodiments of the invention, production of a particular sterol compound may be increased while production of other sterol compounds is decreased. In some embodiments of the invention, production of a plurality of different sterol compounds is increased. In principle, an inventive sterologenic modification may be any chemical, physiological, genetic, or other modification that appropriately alters production of one or more sterol compounds in a host organism produced by that organism as compared with the level produced in an otherwise identical organism not subject to the same modification. In most embodiments, however, the sterologenic modification will comprise a genetic modification, typically resulting in increased production of one or more sterol compounds (e.g., squalene, lanosterol, zymosterol, ergosterol, 7-dehydrocholesterol (provitamin D3) or vitamin D compound(s)). In certain aspects where more than one modification is utilized, such modifications can comprise any combination of chemical, physiological, genetic, or other modification (e.g., one or more genetic modification and chemical or physiological modification).

Ubiquinone biosynthesis polypeptide: The term “ubiquinone biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of ubiquinone. To mention but a few, these ubiquinone biosynthesis polypeptides include, for example, polypeptides of prenyldiphosphate synthase, PHB-polyprenyltransferase, and O-methyltransferase, as well as C₅₋₉ quinone biosynthesis polypeptides. As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, ubiquinone biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other ubiquinone biosynthesis polypeptides.

Ubiquinogenic modification: The term “ubiquinogenic modification”, as used herein, refers to a modification of a host organism that adjusts production of ubiquinone (e.g., CoQ10), as described herein. For example, a ubiquinogenic modification may increase the production level of ubiquinone (e.g., CoQ10), and/or may alter relative levels of ubiquinone and/or ubiquinol. In principle, an inventive ubiquinogenic modification may be any chemical, physiological, genetic, or other modification that appropriately alters production of ubiquinone (e.g., CoQ10) in a host organism produced by that organism as compared with the level produced in an otherwise identical organism not subject to the same modification. In most embodiments, however, the ubiquinogenic modification will comprise a genetic modification, typically resulting in increased production of ubiquinone (CoQ10).

Vitamin D biosynthesis polypeptide: The term “vitamin D biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of one or more vitamin D compounds. To mention but a few, these include, for example, polypeptides enzymes with EC numbers the 1.14.99.7, 5.4.99.7, 5.4.99.8, 5.3.3.5, and/or 1.14.21.6. They further can include the hydroxylases that convert vitamin D₃ to calcitriol (e.g., polypeptides enzymes with EC numbers 1.14.15.- and 1.14.13.13). As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, vitamin D biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other vitamin D biosynthesis polypeptides.

Vitamin E biosynthesis polypeptide: The term “vitamin E biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of vitamin E. To mention but a few, these include, for example, tyrA, pds1(hppd), VTE1, HPT1(VTE2), VTE3, VTE4, and/or GGH polypeptides (i.e., polypeptides that perform the chemical reactions performed by tyrA, pds1(hppd), VTE1, HPT1(VTE2), VTE3, VTE4, and/or GGH, respectively). As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, vitamin E biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other vitamin E biosynthesis polypeptides.

Vitamin K biosynthesis polypeptide: The term “vitamin K biosynthesis polypeptide” refers to any polypeptide that is involved in the synthesis of vitamin K. To mention but a few, these include, for example, MenF, MenD, MenC, MenE, MenB, MenA, UbiE, and/or MenG polypeptides (i.e., polypeptides that perform the chemical reactions performed by MenF, MenD, MenC, MenE, MenB, MenA, UbiE, and/or MenG, respectively). As will be appreciated by those of ordinary skill in the art, in some embodiments of the invention, vitamin K biosynthesis polypeptides include polypeptides that affect the expression and/or activity of one or more other carotenoid biosynthesis polypeptides.

DETAILED DESCRIPTION OF CERTAIN PREFERRED EMBODIMENTS OF THE INVENTION

As noted above, the present invention encompasses the discovery that carotenoids and/or retinolic compounds can desirably be produced in oleaginous yeast and fungi. According to the present invention, strains that both (i) accumulate lipid, often in the form of cytoplasmic oil bodies and typically to at least about 20% of their dry cell weight; and (ii) produce carotenoid(s) and/or retinolic compound(s) at a level at least about 1%, and in some embodiments at least about 3-20%, of their dry cell weight, are generated through manipulation of host cells (i.e., strains, including, e.g., naturally-occurring strains, strains which have been previously modified, etc.). These manipulated host cells are then used to produce carotenoids and/or retinolic compounds, so that carotenoids and/or retinolic compounds that partition into the lipid bodies can readily be isolated.

In general, it will be desirable to balance oleaginy and carotenoid production in inventive cells such that, as soon as a minimum desirable level of oleaginy is achieved, substantially all further carbon which is capable of being utilized and diverted into biosynthesis of products is diverted into a carotenoid and/or retinolic compounds production pathway. In some embodiments of the invention, this strategy involves engineering cells to be oleaginous; in other embodiments, it involves engineering cells to accumulate a higher level of lipid, particularly cytoplasmic lipid, than they would accumulate in the absence of such engineering even though the engineered cells may not become “oleaginous” as defined herein. In other embodiments, the extent to which an oleaginous host cell accumulates lipid is actually reduced so that remaining carbon can be utilized in carotenoid and/or retinolic compound production. According to the present invention, the extent of lipid accumulation in a host cell can be adjusted by modifying the level and/or activity of one or more polypeptides involved in lipid accumulation. Such modification can take the form of genetic engineering and/or exposure to particular growth conditions that induce or inhibit lipid accumulation.

To give but one example of adjustments that could be made to achieve a desired balance between oleaginy and carotenoid and/or retinolic compound production, we note that, while increasing acetyl CoA carboxylase expression (and/or activity) promotes oleaginy, decreasing its expression and/or activity can promote carotenoid and/or retinolic compound production. Those of ordinary skill in the art will appreciate that the expression and/or activity of acetyl CoA carboxylase, or of other polypeptides may be adjusted up or down as desired according to the characteristics of a particular host cell of interest.

We note that engineered cells and processes of using them as described herein may provide one or more advantages as compared with unmodified cells. Such advantages may include, but are not limited to: increased yield (e.g., carotenoid and/or retinolic compound content expressed as either % dry cell weight (mg/mg) or parts per million), titer (g carotenoid/L and/or g retinolic compound/L), specific productivity (mg carotenoid g⁻¹ biomass hour⁻¹ and/or mg retinolic compound g⁻¹ biomass hour⁻¹), and/or volumetric productivity (g carotenoid liter⁻¹ hour⁻¹ and/or g retinolic compound liter⁻¹ hour⁻¹)) of the desired carotenoid and/or retinolic compound (and/or intermediates thereof), and/or decreased formation of undesirable side products (for example, undesirable intermediates).

Thus, for example, the specific productivity for one or more desired carotenoids (e.g. β-carotene, astaxanthin), retinolic compound (e.g., retinol, retinal, retinoic acid), total carotenoids and/or total retinolic compounds may be at or about 0.1, at or about 0.11, at or about 0.12, at or about 0.13, at or about 0.14, at or about 0.15, at or about 0.16, at or about 0.17, at or about 0.18, at or about 0.19, at or about 0.2, at or about 0.21, at or about 0.22, at or about 0.23, at or about 0.24, at or about 0.25, at or about 0.26, at or about 0.27, at or about 0.28, at or about 0.29, at or about 0.3, at or about 0.31, at or about 0.32, at or about 0.33, at or about 0.34, at or about 0.35, at or about 0.36, at or about 0.37, at or about 0.38, at or about 0.39, at or about 0.4, at or about 0.41, at or about 0.42, at or about 0.43, at or about 0.44, at or about 0.45, at or about 0.46, at or about 0.47, at or about 0.48, at or about 0.49, at or about 0.5, at or about 0.51, at or about 0.52, at or about 0.53, at or about 0.54, at or about 0.55, at or about 0.56, at or about 0.57, at or about 0.58, at or about 0.59, at or about 0.6, at or about 0.61, at or about 0.62, at or about 0.63, at or about 0.64, at or about 0.65, at or about 0.66, at or about 0.67, at or about 0.68, at or about 0.69, at or about 0.7, at or about 0.71, at or about 0.72, at or about 0.73, at or about 0.74, at or about 0.75, at or about 0.76, at or about 0.77, at or about 0.78, at or about 0.79, at or about 0.8, at or about 0.81, at or about 0.82, at or about 0.83, at or about 0.84, at or about 0.85, at or about 0.86, at or about 0.87, at or about 0.88, at or about 0.89, at or about 0.9, at or about 0.91, at or about 0.92, at or about 0.93, at or about 0.94, at or about 0.95, at or about 0.96, at or about 0.97, at or about 0.98, at or about 0.99, at or about 1, 1.05, at or about 1.1, at or about 1.15, at or about 1.2, at or about 1.25, at or about 1.3, at or about 1.35, at or about 1.4, at or about 1.45, at or about 1.5, at or about 1.55, at or about 1.6, at or about 1.65, at or about 1.7, at or about 1.75, at or about 1.8, at or about 1.85, at or about 1.9, at or about 1.95, at or about 2 mg g⁻¹ hour⁻¹ or more.

Thus, for example, the volumetric productivity for one or more desired carotenoids (e.g. β-carotenoid, astaxanthin), retinolic compound (e.g., retinol, retinal, retinoic acid), total carotenoids and/or total retinolic compounds may be at or about 0.01, at or about 0.011, at or about 0.012, at or about 0.013, at or about 0.014, at or about 0.015, at or about 0.016, at or about 0.017, at or about 0.018, at or about 0.019, at or about 0.02, at or about 0.021, at or about 0.022, at or about 0.023, at or about 0.024, at or about 0.025, at or about 0.026, at or about 0.027, at or about 0.028, at or about 0.029, at or about 0.03, at or about 0.031, at or about 0.032, at or about 0.033, at or about 0.034, at or about 0.035, at or about 0.036, at or about 0.037, at or about 0.038, at or about 0.039, at or about 0.04, at or about 0.041, at or about 0.042, at or about 0.043, at or about 0.044, at or about 0.045, at or about 0.046, at or about 0.047, at or about 0.048, at or about 0.049, at or about 0.05, at or about 0.051, at or about 0.052, at or about 0.053, at or about 0.054, at or about 0.055, at or about 0.056, at or about 0.057, at or about 0.058, at or about 0.059, at or about 0.06, at or about 0.061, at or about 0.062, at or about 0.063, at or about 0.064, at or about 0.065, at or about 0.066, at or about 0.067, at or about 0.068, at or about 0.069, at or about 0.07, at or about 0.071, at or about 0.072, at or about 0.073, at or about 0.074, at or about 0.075, at or about 0.076, at or about 0.077, at or about 0.078, at or about 0.079, at or about 0.08, at or about 0.081, at or about 0.082, at or about 0.083, at or about 0.084, at or about 0.085, at or about 0.086, at or about 0.087, at or about 0.088, at or about 0.089, at or about 0.09, at or about 0.091, at or about 0.092, at or about 0.093, at or about 0.094, at or about 0.095, at or about 0.096, at or about 0.097, at or about 0.098, at or about 0.099, at or about 0.1, 0.105, at or about 0.110, at or about 0.115, at or about 0.120, at or about 0.125, at or about 0.130, at or about 0.135, at or about 0.14, at or about 0.145, at or about 0.15, at or about 0.155, at or about 0.16, at or about 0.165, at or about 0.17, at or about 0.175, at or about 0.18, at or about 0.185, at or about 0.19, at or about 0.195, at or about 0.20 grams liter⁻¹ hour⁻¹ or more.

Host Cells

Those of ordinary skill in the art will readily appreciate that a variety of yeast and fungal strains exist that are naturally oleaginous or that naturally produce carotenoids. Yeast and fungal strains do not naturally produce retinolic compounds. Any of such strains may be utilized as host strains according to the present invention, and may be engineered or otherwise manipulated to generate inventive oleaginous, carotenoid-producing strains and/or oleaginous, retinolic acid compound-producing strains. Alternatively, strains that naturally are neither oleaginous nor: i) carotenoid-producing and/or ii) retinolic compound-producing may be employed. Furthermore, even when a particular strain has a natural capacity for oleaginy or for carotenoid production, its natural capabilities may be adjusted as described herein, so as to change the production level of lipid, carotenoid and/or retinolic compound. In certain embodiments engineering or manipulation of a strain results in modification of a type of lipid, carotenoid and/or retinolic compound which is produced. For example, a strain may be naturally oleaginous and/or carotenogenic, however engineering or modification of the strain may be employed so as to change the type of lipid which is accumulated and or to change the type of carotenoid which is produced. Additionally or alternatively, naturally oleaginous strain may be engineered to permit retinolic compound production. Moreover, further engineering or modification of the strain may be employed so as to change the type of lipid which is accumulated and/or to change the type of retinolic compound which is produced.

When selecting a particular yeast or fungal strain for use in accordance with the present invention, it will generally be desirable to select one whose cultivation characteristics are amenable to commercial scale production. For example, it will generally (though not necessarily always) be desirable to avoid filamentous organisms, or organisms with particularly unusual or stringent requirements for growth conditions. However, where conditions for commercial scale production can be applied which allow for utilization of filamentous organisms, these may be selected as host cells. In some embodiments of the invention, it will be desirable to utilize edible organisms as host cells, as they may optionally be formulated directly into food or feed additives, or into nutritional supplements, as desired. For ease of production, some embodiments of the invention utilize host cells that are genetically tractable, amenable to molecular genetics (e.g., can be efficiently transformed, especially with established or available vectors; optionally can incorporate and/or integrate multiple genes, for example sequentially; and/or have known genetic sequence; etc), devoid of complex growth requirements (e.g., a necessity for light), mesophilic (e.g., prefer growth temperatures with in the range of about 20-32° C.) (e.g. 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32° C.), able to assimilate a variety of carbon and nitrogen sources and/or capable of growing to high cell density. Alternatively or additionally, various embodiments of the invention utilize host cells that grow as single cells rather than multicellular organisms (e.g., as mycelia).

In general, when it is desirable to utilize a naturally oleaginous organism in accordance with the present invention, any modifiable and cultivatable oleaginous organism may be employed. In certain embodiments of the invention, yeast or fungi of genera including, but not limited to, Blakeslea, Candida, Cryptococcus, Cunninghamella, Lipomyces, Mortierella, Mucor, Phycomyces, Pythium, Rhodosporidium, Rhodotorula, Trichosporon, and Yarrowia are employed. In certain particular embodiments, organisms of species that include, but are not limited to, Blakeslea trispora, Candida pulcherrima, C. revkaufi, C. tropicalis, Cryptococcus curvatus, Cunninghamella echinulata, C. elegans, C. japonica, Lipomyces starkeyi, L. lipoferus, Mortierella alpina, M. isabellina, M. ramanniana, M. vinacea, Mucor circinelloides, Phycomyces blakesleanus, Pythium irregulare, Rhodosporidium toruloides, Rhodotorula glutinis, R. gracilis, R. graminis, R. mucilaginosa, R. pinicola, Trichosporon pullans, T. cutaneum, and Yarrowia lipolytica are used.

Of these naturally oleaginous strains, some also naturally produce carotenoids and some do not; these strains do not naturally produced retinolic compounds. In most cases, only low levels (less than about 0.05% dry cell weight) of carotenoids are produced by naturally-occurring carotenogenic, oleaginous yeast or fungi. Higher levels of β-carotene are sometimes produced, but high levels of other carotenoids are generally not observed.

In general, any organism that is naturally oleaginous and non-carotenoid-producing (e.g., produce less than about 0.05% dry cell weight, do not produce the carotenoid of interest) may be utilized as a host cell in accordance with the present invention. Additionally or alternatively, any organism that is naturally oleaginous and non-retinolic compound-producing (e.g., produce less than about 0.05% dry cell weight, do not produce the retinolic compound of interest) may be utilized as a host cell in accordance with the present invention. For example, introduction of one or more retinologenic modifications (e.g., increased expression of one or more endogenous or heterologous retinologenic polypeptides), in accordance with the present invention, can achieve the goals for retinolic compound production. In some embodiments, the organism is a yeast or fungus from a genus such as, but not limited to, Candida, Cryptococcus, Cunninghamella, Lipomyces, Mortierella, Pythium, Trichosporon, and Yarrowia; in some embodiments, the organism is of a species including, but not limited to, Mortierella alpina and Yarrowia lipolytica.

Comparably, the present invention may utilize any naturally oleaginous, carotenoid-producing organism as a host cell. In general, the present invention may be utilized to increase carbon flow into the isoprenoid pathway in naturally carotenoid-producing organisms (particularly for organisms other than Blakeslea and Phycomyces), and/or to shift production from one carotenoid (e.g., β-carotene) to another (e.g., astaxanthin). Introduction of one or more carotenogenic modifications (e.g., increased expression of one or more endogenous or heterologous carotenogenic polypeptides), in accordance with the present invention, can achieve these goals. Additionally or alternatively, the present invention may be utilized to introduce the ability to produce one or more retinolic compounds in such naturally carotenoid-producing host cells.

In certain embodiments of the invention, the utilized oleaginous, carotenoid-producing organism is a yeast or fungus, for example of a genus such as, but not limited to, Blakeslea, Mucor, Phycomyces, Rhodosporidium, and Rhodotorula; in some embodiments, the organism is of a species such as, Mucor circinelloides and Rhodotorula glutinis.

When it is desirable to utilize strains that are naturally non-oleaginous as host cells in accordance with the present invention, genera of non-oleaginous yeast or fungi include, but are not limited to, Aspergillus, Botrytis, Cercospora, Fusarium (Gibberella), Kluyveromyces, Neurospora, Penicillium, Pichia (Hansenula), Puccinia, Saccharomyces, Sclerotium, Trichoderma, and Xanthophyllomyces (Phaffia); in some embodiments, the organism is of a species including, but not limited to, Candida utilis, Aspergillus nidulans, A. niger, A. terreus, Botrytis cinerea, Cercospora nicotianae, Fusarium fujikuroi (Gibberella zeae), Kluyveromyces lactis, K. lactis, Neurospora crassa, Pichia pastoris, Puccinia distincta, Saccharomyces cerevisiae, Sclerotium rolfsii, Trichoderma reesei, and Xanthophyllomyces dendrorhous (Phaffia rhodozyma).

It will be appreciated that the term “non-oleaginous”, as used herein, encompasses both strains that naturally have some ability to accumulate lipid, especially cytoplasmically, but do not do so to a level sufficient to qualify as “oleaginous” as defined herein, as well as strains that do not naturally have any ability to accumulate extra lipid, e.g., extra-membranous lipid. It will further be appreciated that, in some embodiments of the invention, it will be sufficient to increase the natural level of oleaginy of a particular host cell, even if the modified cell does not qualify as oleaginous as defined herein. In some embodiments, the cell will be modified to accumulate at least about 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, or 5% in dry cell weight as lipid, so long as the accumulation level is more than that observed in the unmodified parental cell.

As with the naturally oleaginous organisms, some of the naturally non-oleaginous fungi naturally produce carotenoids, whereas others do not; these strains do not naturally produced retinolic compounds. Genera of naturally non-oleaginous fungi that do not naturally produce carotenoids (e.g., produce less than about 0.05% dry cell weight, do not produce a carotenoid or retinolic compound of interest) may desirably be used as host cells in accordance with the present invention include, but are not limited to, Aspergillus, Kluyveromyces, Penicillium, Saccharomyces, and Pichia; species include, but are not limited to, Candida utilis, Aspergillus niger and Saccharomyces cerevisiae. Genera of naturally non-oleaginous fungi that do naturally produce carotenoids or retinolic compounds and that may desirably be used as host cells in accordance with the present invention include, but are not limited to, Botrytis, Cercospora, Fusarium (Gibberella), Neurospora, Puccinia, Sclerotium, Trichoderma, and Xanthophyllomyces (Phaffia); species include, but are not limited to, Xanthophyllomyces dendrorhous (Phaffia rhodozyma).

As discussed above, any of a variety of organisms may be employed as host cells in accordance with the present invention. In certain embodiments of the invention, host cells will be Yarrowia lipolytica cells. Advantages of Y. lipolytica include, for example, tractable genetics and molecular biology, availability of genomic sequence (see, for example. Sherman et al. Nucleic Acids Res. 32 (Database issue):D315-8, 2004), suitability to various cost-effective growth conditions, and ability to grow to high cell density. In addition, Y. lipolytica is naturally oleaginous, such that fewer manipulations may be required to generate an oleaginous, carotenoid-producing and/or retinolic compound-producing Y. lipolytica strain than might be required for other organisms. Furthermore, there is already extensive commercial experience with Y. lipolytica.

Saccharomyces cerevisiae is also a useful host cell in accordance with the present invention, particularly due to its experimental tractability and the extensive experience that researchers have accumulated with the organism. Although cultivation of Saccharomyces under high carbon conditions may result in increased ethanol production, this can generally be managed by process and/or genetic alterations.

Additional useful hosts include Xanthophyllomyces dendrorhous (Phaffia rhodozyma), which is experimentally tractable and naturally carotenogenic. Xanthophyllomyces dendrorhous (Phaffia rhodozyma) strains can produce several carotenoids, including astaxanthin.

Aspergillus niger and Mortierella alpina accumulate large amounts of citric acid and fatty acid, respectively; Mortierella alpina is also oleaginous.

Neurospora or Gibberella are also useful. They are not naturally oleaginous and tend to produce very low levels of carotenoids, thus extensive modification may be required in accordance with the present invention. Neurospora and Gibberella are considered relatively tractable from an experimental standpoint. Both are filamentous fungi, such that production at commercial scales can be a challenge necessary to overcome in utilization of such strains.

Mucor circinelloides is another available useful species. While its molecular genetics are generally less accessible than are those of some other organisms, it naturally produces β-carotene, thus may require less modification than other species available.

Candida utilis is a further useful species. Although it is not naturally oleaginous and produces little or no carotenoids, it is amenable to genetic manipulation (for example, see Iwakiri et al. (2006) Yeast 23:23-34, Iwakiri et al. (2005) Yeast 2005 22:1079-87, Iwakiri et al. (2005) Yeast 22:1049-60, Rodriquez et al. (1998) Yeast 14:1399-406, Rodriquez et al. (1998) FEMS Microbiol Lett. 165:335-40, and Kondo et al. (1995) J Bacteriol. 177:7171-7) and furthermore is edible.

Molecular genetics can be performed in Blakeslea, though significant effort may be required. Furthermore, cost-effective fermentation conditions can be challenging, as, for example, it may be required that the two mating types are mixed. Fungi of the genus Phycomyces are also possible sources which have the potential to pose fermentation process challenges, and these fungi may be less amenable to manipulate than several other potential host organisms.

Additional useful hosts include strains such as Schizosaccharomyces pombe, Saitoella complicata, and Sporidiobolus ruineniae.

Those of ordinary skill in the art will appreciate that the selection of a particular host cell for use in accordance with the present invention will also affect, for example, the selection of expression sequences utilized with any heterologous polypeptide to be introduced into the cell, codon bias that can optionally be engineered into any nucleic acid to be expressed in the cell, and will also influence various aspects of culture conditions, etc. Much is known about the different gene regulatory requirements, protein targeting sequence requirements, and cultivation requirements, of different host cells to be utilized in accordance with the present invention (see, for example, with respect to Yarrowia, Barth et al. FEMS Microbiol Rev. 19:219, 1997; Madzak et al. J Biotechnol. 109:63, 2004; see, for example, with respect to Xanthophyllomyces, Verdoes et al. Appl Environ Microbiol 69: 3728-38, 2003; Visser et al. FEMS Yeast Res 4: 221-31, 2003; Martinez et al. Antonie Van Leeuwenhoek. 73(2):147-53, 1998; Kim et al. Appl Environ Microbiol. 64(5):1947-9, 1998; Wery et al. Gene. 184(1):89-97, 1997; see, for example, with respect to Saccharomyces, Guthrie and Fink Methods in Enzymology 194:1-933, 1991). In certain aspects, for example, targeting sequences of the host cell (or closely related analogs) may be useful to include for directing heterologous proteins to subcellular localization. Thus, such useful targeting sequences can be added to heterologous sequence for proper intracellular localization of activity. In other aspects (e.g., addition of mitochondrial targeting sequences), heterologous targeting sequences may be eliminated or altered in the selected heterologous sequence (e.g., alteration or removal of source organism plant chloroplast targeting sequences).

To give but a few specific examples, of promoters and/or regulatory sequences that may be employed in expression of polypeptides according to the present invention, useful promoters include, but are not limited to, the Leu2 promoter and variants thereof (see, for example, see U.S. Pat. No. 5,786,212); the EF1alpha protein and ribosomal protein S7 gene promoters (see, for example, PCT Application WO 97/44470); the Gpm (see US20050014270), Xpr2 (see U.S. Pat. No. 4,937,189), Tef1, Gpd1 (see, for example, US Application 2005-0014270A1), Cam1 (YALI0C24420g), YALI0D16467g, Tef4 (YALI0B12562g), Yef3 (YALI0E13277g), Pox2, Yat1 (see, for example US Application 2005-0130280; PCT Application WO 06/052754), Fba1 (see, for example WO05049805), and/or Gpat (see WO06031937) promoters; the sequences represented by SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, subsequences thereof, and hybrid and tandem derivatives thereof (e.g., as disclosed in US Application 2004-0146975); the sequences represented by SEQ ID NO: 1, 2, or 3 including fragments (e.g. by 462-1016 and by 197-1016 of SEQ ID NO: 1; by 5-523 of SEQ ID NO:3) and complements thereof (e.g., as disclosed in US patent number http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO1&Sect2=HITOFF&d=PALL&p=1&u=%2Fnetahtml%2FPTO%2Fsrchnum.htm&r=1&f=G&1=50&s1=5952195.PN.&OS=PN/5952195&RS=PN/5952195-h0#h0http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO1&Sect2=HITOFF&d=PALL&p=1&u=%2Fnetahtml%2FPTO%2Fsrchnum.htm&r=1&f=G&1=50&s1=5952195.PN.&OS=PN/5952195&RS=PN/5952195-h2#h25,952,195); CYP52A2A (see, for example, US Application 2002-0034788); promoter sequences from fungal (e.g., C. tropicalis) catalase, citrate synthase, 3-ketoacyl-CoA thiolase A, citrate synthase, O-acetylhornserine sulphydrylase, protease, carnitine O-acetyltransferase, hydratase-dehydrogenase, epimerase genes; promoter sequences from Pox4 genes (see, for example, US application 2004-0265980); and/or promoter sequences from Met2, Met3, Met6, Met25 and YALI0D12903g genes. Any such promoters can be used in conjunction with endogenous genes and/or heterologous genes for modification of expression patterns of endogenous polypeptides and/or heterologous polypeptides in accordance with the present invention.

Alternatively or additionally, regulatory sequences useful in accordance with the present invention may include one or more Xpr2 promoter fragments, for example as described in US patent http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO1&Sect2=HITOFF&d=PALL&p=1&u=%2Fnetahtml%2FPTO%2Fsrchnum.htm&r=1&f=G&1=50&s1=6083717.PN.&OS=PN/6083717&RS=PN/6083717-h0#h0http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO1&Sect2=HITOFF&d=PALL&p=1&u=%2Fnetahtml%2FPTO%2Fsrchnum.htm&r=1&f=G&1=50&s1=6083717.PN.&OS=PN/6083717&RS=PN/6083717-h2#h26,083,717 (e.g. SEQ ID NOS: 1-4 also including sequences with 80% or more identity to these SEQ ID NOs) (e.g., see Example 11) in one or more copies either in single or in tandem. Similarly, exemplary terminator sequences include, but are not limited to, Y. lipolytica Xpr2 (see U.S. Pat. No. 4,937,189) and Pox2 (YALI0F10857g) terminator sequences.

In some embodiments of this invention, it may be desirable to fused sequences encoding specific targeting signals to bacterial source genes. For example, in certain embodiments mitochondrial signal sequences are useful in conjunction with, e.g., bacterial polypeptides for effective targeting of polypeptides for proper functioning. Mitochondrial signal sequences are known in the art, and include, but are not limited to example, mitochondrial signal sequences provided in Table 52 below. In other embodiments, it may be desirable to utilize genes from other source organisms such as animals, plants, alga, or microalgae, fungi, yeast, insect, protozoa, and mammals.

TABLE 52 Examples of mitochondrial targeting sequences. Protein Species (residues) GI Sequence Yarrowia NUAM 6689648 MLSRNLSKFARAGLIRPATTSTHTRLFSVSARRL lipoylitica (AA 1-34) (SEQ ID NO: 171) Yarrowia NUHM 50549567 MLRLIRPRLAALARPTTRAPQALNARTHIVSV lipoylitica (AA 1-32) (SEQ ID NO: 172) Saccharomyces Coq1 536190 MFQRSGAAHHIKLISSRRCRFKSSFAVALNAA cerevisiae (AA 1-53) SKLVTPKILWNNPISLVSKEM  (SEQ ID NO: 173) Yarrowia Coq1 60389562 MLRVGRIGTKTLASSSLRFVAGARPKSTLTEA lipoylitica (AA 1-77) VLETTGLLKTTPQNPEWSGAVKQASRLVETD TPIRDPFSIVSQEM (SEQ ID NO: 174) Engineering Oleaginy

All living organisms synthesize lipids for use in their membranes and various other structures. However, most organisms do not accumulate in excess of about 10% of their dry cell weight as total lipid, and most of this lipid generally resides within cellular membranes.

Significant biochemical work has been done to define the metabolic enzymes necessary to confer oleaginy on microorganisms (primarily for the purpose of engineering single cell oils as commercial sources of arachidonic acid and docosahexaenoic acid; see for example Ratledge Biochimie 86:807, 2004, the entire contents of which are incorporated herein by reference). Although this biochemical work is compelling, there only have been a limited number of reports of de novo oleaginy being established through genetic engineering with the genes encoding the key metabolic enzymes. It should be noted that oleaginous organisms typically accumulate lipid only when grown under conditions of carbon excess and nitrogen limitation. The present invention further establishes that the limitation of other nutrients (e.g. phosphate and/or magnesium) can also induce lipid accumulation. The present invention establishes, for example, that limitation of nutrients such as phosphate and/or magnesium can induce lipid accumulation, much as is observed under conditions of nitrogen limitation. Under these conditions, the organism readily depletes the limiting nutrient but continues to assimilate the carbon source. The “excess” carbon is channeled into lipid biosynthesis so that lipids (usually triacylglycerols) accumulate in the cytosol, typically in the form of bodies. It should be noted that oleaginous organisms typically only accumulate lipid when grown under conditions of carbon excess and nitrogen or other nutrient limitation (e.g. phosphate or magnesium). Under these conditions, the organism readily depletes the limiting nutrient but continues to assimilate the carbon source. The “excess” carbon is channeled into lipid biosynthesis so that lipids (usually triacylglycerols) accumulate in the cytosol, typically in the form of bodies.

In general, it is thought that, in order to be oleaginous, an organism must produce both acetyl-CoA and NADPH in the cytosol, which can then be utilized by the fatty acid synthase machinery to generate lipids. In at least some oleaginous organisms, acetyl-CoA is generated in the cytosol through the action of ATP-citrate lyase, which catalyzes the reaction: citrate+CoA+ATP→acetyl-CoA+oxaloacetate+ADP+P_(i).  (1)

Of course, in order for ATP-citrate lyase to generate appropriate levels of acetyl-CoA in the cytosol, it must first have an available pool of its substrate citric acid. Citric acid is generated in the mitochondria of all eukaryotic cells through the tricarboxylic acid (TCA) cycle, and can be moved into the cytosol (in exchange for malate) by citrate/malate translocase.

In most oleaginous organisms, and in some non-oleaginous organisms, the enzyme isocitrate dehydrogenase, which operates as part of the TCA cycle in the mitochondria, is strongly AMP-dependent. Thus, when AMP is depleted from the mitochondria, this enzyme is inactivated. When isocitrate dehydrogenase is inactive, isocitrate accumulates in the mitochondria. This accumulated isocitrate is then equilibrated with citric acid, presumably through the action of aconitase. Therefore, under conditions of low AMP, citrate accumulates in the mitochondria. As noted above, mitochondrial citrate is readily transported into the cytosol.

AMP depletion, which in oleaginous organisms is believed to initiate the cascade leading to accumulation of citrate (and therefore acetyl-CoA) in the cytoplasm, occurs as a result of the nutrient depletion mentioned above. When oleaginous cells are grown in the presence of excess carbon source but under conditions limiting for nitrogen or some other nutrient(s) (e.g., phosphate or magnesium), the activity of AMP deaminase, which catalyzes the reaction: AMP→inosine 5′-monophosphate+NH₃  (2) is strongly induced. The increased activity of this enzyme depletes cellular AMP in both the cytosol and the mitochondria. Depletion of AMP from the mitochondria is thought to inactivate the AMP-dependent isocitrate dehydrogenase, resulting in accumulation of citrate in the mitochondria and, therefore, the cytosol. This series of events is depicted diagrammatically in FIG. 2.

As noted above, oleaginy requires both cytosolic acetyl-CoA and cytosolic NADPH. It is believed that, in many oleaginous organisms, appropriate levels of cytosolic NADPH are provided through the action of malic enzyme (Enzyme 3 in FIG. 2). Some oleaginous organisms (e.g., Lipomyces and some Candida) do not appear to have malic enzymes, however, so apparently other enzymes can provide comparable activity, although it is expected that a dedicated source of NADPH is probably required for fatty acid synthesis (see, for example, Wynn et al., Microbiol 145:1911, 1999; Ratledge Adv. Appl. Microbiol. 51:1, 2002, each of which is incorporated herein by reference in its entirety).

Other activities which can be involved in regenerating NADPH include, for example, 6-phosphogluconate dehydrogenase (gnd); Fructose 1,6 bisphosphatase (fbp); Glucose 6 phosphate dehydrogenase (g6pd); NADH kinase (EC 2.7.1.86); and/or transhydrogenase (EC 1.6.1.1 and 1.6.1.2).

Gnd is part of the pentose phosphate pathway and catalyses the reaction: 6-phospho-D-gluconate+NADP+→D-ribulose 5-phosphate+CO₂+NADPH

-   Fbp is a hydrolase that catalyses the gluconeogenic reaction:     D-fructose 1,6-bisphosphate+H₂O→D-fructose 6-phosphate+phosphate     Fbp redirects carbon flow from glycolysis towards the pentose     phosphate pathway. The oxidative portion of the pentose phosphate     pathway, which includes glucose 6 phosphate dehydrogenase and     6-phosphogluconate dehydrogenase, enables the regeneration of NADPH.     G6pd is part of the pentose phosphate pathway and catalyses the     reaction:     D-glucose 6-phosphate+NADP⁺→D-glucono-1,5-lactone     6-phosphate+NADPH+H⁺NADH -   kinase catalyzes the reaction:     ATP+NADH→ADP+NADPH

Transhydrogenase catalyzes the reaction: NADPH+NAD⁺⇄NADP⁺+NADH Thus, enhancing the expression and/or activity of any of these enzymes can increase NADPH levels and promote anabolic pathways requiring NADPH.

Alternative or additional strategies to promote oleaginy may include one or more of the following: (1) increased or heterologous expression of one or more of acyl-CoA:diacylglycerol acyltransferase (e.g., DGA1; YALI0E32769g); phospholipid:diacylglycerol acyltransferase (e.g., LRO1; YALI0E16797g); and acyl-CoA:cholesterol acyltransferase (e.g., ARE genes such as ARE1, ARE2, YALI0F06578g), which are involved in triglyceride synthesis (Kalscheuer et al. Appl Environ Microbiol p. 7119-7125, 2004; Oelkers et al. J Biol Chem 277:8877-8881, 2002; and Sorger et al. J Biol Chem 279:31190-31196, 2004), (2) decreased expression of triglyceride lipases (e.g., TGL3 and/or TGL4; YALI0D17534g and/or YALI0F10010g (Kurat et al. J Biol Chem 281:491-500, 2006); and (3) decreased expression of one or more acyl-coenzyme A oxidase activities, for example encoded by POX genes (e.g. POX1, POX2, POX3, POX4, POX5; YALI0C23859g, YALI0D24750g, YALI0E06567g, YALI0E27654g, YALI0E32835g, YALI0F10857g; see for example Mlickova et al. Appl Environ Microbiol 70: 3918-3924, 2004; Binns et al. J Cell Biol 173:719, 2006).

Thus, according to the present invention, the oleaginy of a host organism may be enhanced by modifying the expression or activity of one or more polypeptides involved in generating cytosolic acetyl-CoA and/or NADPH and/or altering lipid levels through other mechanisms. For example, modification of the expression or activity of one or more of acetyl-CoA carboxylase, pyruvate decarboxylase, isocitrate dehydrogenase, ATP-citrate lyase, malic enzyme, AMP-deaminase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, fructose 1,6 bisphosphatase, NADH kinase, transhydrogenase, acyl-CoA:diacylglycerol acyltransferase, phospholipid:diacylglycerol acyltransferase, acyl-CoA:cholesterol acyltransferase, triglyceride lipase, acyl-coenzyme A oxidase can enhance oleaginy in accordance with the present invention. Exemplary polypeptides which can be utilized or derived so as to enhance oleaginy in accordance with the present invention include, but are not limited to those acetyl-CoA carboxylase, pyruvate decarboxylase, isocitrate dehydrogenase, ATP-citrate lyase, malic enzyme, AMP-deaminase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, fructose 1,6 bisphosphatase, NADH kinase, transhydrogenase, acyl-CoA:diacylglycerol acyltransferase, phospholipid:diacylglycerol acyltransferase, acyl-CoA:cholesterol acyltransferase, triglyceride lipase, acyl-coenzyme A oxidase polypeptides provided in Tables 1-6, and 31-47, respectively.

In some embodiments of the invention, where an oleaginous host cell is employed, enzymes and regulatory components relevant to oleaginy are already in place but could be modified, if desired, by for example altering expression or activity of one or more oleaginic polypeptides and/or by introducing one or more heterologous oleaginic polypeptides. In those embodiments of the invention where a non-oleaginous host cell is employed, it is generally expected that at least one or more heterologous oleaginic polypeptides will be introduced.

The present invention contemplates not only introduction of heterologous oleaginous polypeptides, but also adjustment of expression or activity levels of heterologous or endogenous oleaginic polypeptides, including, for example, alteration of constitutive or inducible expression patterns. In some embodiments of the invention, expression patterns are adjusted such that growth in nutrient-limiting conditions is not required to induce oleaginy. For example, genetic modifications comprising alteration and/or addition of regulatory sequences (e.g., promoter elements, terminator elements) and/or regulatory factors (e.g., polypeptides that modulate transcription, splicing, translation, modification, etc.) may be utilized to confer particular regulation of expression patterns. Such genetic modifications may be utilized in conjunction with endogenous genes (e.g., for regulation of endogenous oleaginic polypeptide(s)); alternatively, such genetic modifications may be included so as to confer regulation of expression of at least one heterologous polypeptide (e.g., oleaginic polypeptide(s)).

In some embodiments, at least one oleaginic polypeptide is introduced into a host cell. In some embodiments of the invention, a plurality (e.g., two or more) of different oleaginic polypeptides is introduced into the same host cell. In some embodiments, the plurality of oleaginic polypeptides contains polypeptides from the same source organism; in other embodiments, the plurality includes polypeptides independently selected from different source organisms.

Representative examples of a variety of oleaginic polypeptides that may be introduced into or modified within host cells according to the present invention, include, but are not limited to, those provided in Tables 1-6, and Tables 31-47. As noted above, it is expected that at least some of these polypeptides (e.g., malic enzyme and ATP-citrate lyase) should desirably act in concert, and possibly together with one or more components of fatty acid synthase, such that, in some embodiments of the invention, it will be desirable to utilize two or more oleaginic polypeptides from the same source organism.

In certain embodiments, the oleaginy of a host organism is enhanced by growing the organism on a carbon source comprising one or more oils. For example, an organism may be grown on a carbon source comprising one or more oils selected from the group consisting of, for example, olive, canola, corn, sunflower, soybean, cottonseed, rapeseed, etc., and combinations thereof. In certain embodiments, the oleaginy of a host organism is enhanced by growing the organism on a carbon source comprising one or more oils in combination with modifying the expression or activity of one or more polypeptides such as any of those described above (e.g., oleaginic polypeptides such as polypeptides involved in generating cytosolic acetyl-CoA and/or NADPH) and/or altering lipid levels through other mechanisms.

In general, source organisms for oleaginic polypeptides to be used in accordance with the present invention include, but are not limited to, Blakeslea, Candida, Cryptococcus, Cunninghamella, Lipomyces, Mortierella, Mucor, Phycomyces, Pythium, Rhodosporidium, Rhodotorula, Trichosporon, Yarrowia, Aspergillus, Botrytis, Cercospora, Fusarium (Gibberella), Kluyveromyces, Neurospora, Penicillium, Pichia (Hansenula), Puccinia, Saccharomyces, Sclerotium, Trichoderma, and Xanthophyllomyces (Phaffia). In some embodiments, the source species for acetyl CoA carboxylase, ATP-citrate lyase, malice enzyme and/or AMP deaminase polypeptides include, but are not limited to, Aspergillus nidulans, Cryptococcus neoformans, Fusarium fujikuroi, Kluyveromyces lactis, Neurospora crassa, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Ustilago maydis, and Yarrowia lipolytica; in some embodiments, source species for pyruvate decarboxylase or isocitrate dehydrogenase polypeptides include, but are not limited to Neurospora crassa, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), Aspergillus niger, Saccharomyces cerevisiae, Mucor circinelloides, Rhodotorula glutinis, Candida utilis, Mortierella alpina, and Yarrowia lipolytica.

Aspergillus niger accumulates large amounts of citric acid, whereas Mortierella alpina and Thraustochytrium sp. accumulate large amounts of fatty acid, respectively; Mortierella alpina and Thraustochytrium are also oleaginous.

To give but one particular example of a host cell engineered to be oleaginous (or at least to accumulate increased levels of lipid) in accordance with the present invention, S. cerevisiae can be engineered to express one or more oleaginic polypeptides, e.g., from heterologous source organisms. In some embodiments, a plurality of different oleaginic polypeptides are expressed, optionally from different source organisms. For instance, in some embodiments, S. cerevisiae cells are engineered to express (and/or to increase expression of) ATP-citrate lyase (e.g., from N. crassa), AMP deaminase (e.g., from S. cerevisiae), and/or malic enzyme (e.g., from M. circinelloides). In other embodiments, Candida utilis and Phaffia rhodozyma can be similarly modified. Modified S. cerevisiae, C. utilis, and P. rhodozyma strains can be further modified as described herein to increase production of one or more carotenoids.

In certain embodiments, host cells are engineered to be oleaginous by introducing one or more oleaginic polypeptides. In general, any oleaginic polypeptide can be introduced into any host cell of the present invention. In certain embodiments, such oleaginic polypeptides are codon-optimized to accommodate the codon preferences of the host cell. In certain embodiments, an oleaginic polypeptide introduced into a host cell is from the same organism as the host cell and/or a related organism. For example, without limitation, the present invention encompasses the recognition that it may be desirable to introduce a fungal oleaginic polypeptide into a fungal host cell (e.g., from the same and/or a related fungal species). In certain embodiments, the host cell is a Y. lipolytica host cell. In certain aspects of such embodiments, a Y. lipolytica oleaginic polypeptide is introduced into the Y. lipolytica host cell. In certain aspects, a S. cerevisiae oleaginic polypeptide is introduced into the Y. lipolytica host cell. In certain aspects, any of a variety of fungal oleaginic polypeptides is introduced into the Y. lipolytica host cell.

Engineering Carotenoid Production

Carotenoids are synthesized from isoprenoid precursors, some of which are also involved in the production of steroids and sterols. The most common isoprenoid biosynthesis pathway, sometimes referred to as the “mevalonate pathway”, is generally depicted in FIG. 3. As shown, acetyl-CoA is converted, via hydroxymethylglutaryl-CoA (HMG-CoA), into mevalonate. Mevalonate is then phosphorylated and converted into the five-carbon compound isopentenyl pyrophosphate (IPP). Following isomerization of IPP into dimethylallyl pyrophosphate (DMAPP), three sequential condensation reactions with additional molecules of IPP generate the ten-carbon molecule geranyl pyrophosphate (GPP), followed by the fifteen-carbon molecule farnesyl pyrophosphate (FPP), and finally the twenty-carbon compound geranylgeranyl pyrophosphate (GGPP).

An alternative isoprenoid biosynthesis pathway, that is utilized by some organisms (particularly bacteria) and is sometimes called the “mevalonate-independent pathway”, is depicted in FIG. 4. This pathway is initiated by the synthesis of 1-deoxy-D-xyloglucose-5-phosphate (DOXP) from pyruvate and glyceraldehyde-3-phosphate. DOXP is then converted, via a series of reactions shown in FIG. 4, into IPP, which isomerizes into DMAPP and is then converted, via GPP and FPP, into GGPP as shown in FIG. 3 and discussed above.

Various proteins involved in isoprenoid biosynthesis have been identified and characterized in a number of organisms. Moreover, various aspects of the isoprenoid biosynthesis pathway are conserved throughout the fungal, bacterial, plant and animal kingdoms. For example, polypeptides corresponding to the acetoacetyl-CoA thiolase, HMG-CoA synthase, HMG-CoA reductase, mevalonate kinase, phosphomevalonate kinase, mevalonate pyrophosphate decarboxylase, IPP isomerase, FPP synthase, and GGPP synthase shown in FIG. 3 have been identified in and isolated from a wide variety of organisms and cells. Representative examples of a wide variety of such polypeptides are provided in Tables 7-15. One or more of the polypeptides selected from those provided in any one of Tables 7-15 may be utilized or derived for use in the methods and compositions in accordance with the present invention.

Alternatively or additionally, modified mevalonate kinase polypeptides that exhibit decreased feedback inhibition properties (e.g. to farnesyl pyrophosphate (FPP)) may be utilized in accordance with the present invention. Such modified mevalonate kinase polypeptides may be of eukaryotic or prokaryotic origin. For example, modified versions of mevalonate kinase polypeptides from animals (including humans), plants, algae, fungi (including yeast), and/or bacteria may be employed; for instance, modified versions of mevalonate kinase polypeptides disclosed in Table 10 herein may be utilized.

Particular examples of modified mevalonate kinase polypeptides include “feedback-resistant mevalonate kinases” disclosed in PCT Application WO 2006/063752. Thus, for example, a modified mevalonate kinase polypeptide may include one or more mutation (s) at one or more amino acid position (s) selected from the group consisting of amino acid positions corresponding to positions 17, 47, 93, 94, 132, 167, 169, 204, and 266 of the amino acid sequence of Paracoccus zeaxanthinifaciens mevalonate kinase as shown in SEQ ID NO: 1 of PCT Application WO 2004/111214. For example, the modified mevalonate kinase polypeptide may contain one or more substitutions at positions corresponding to one or more of I17T, G47D, K93E, V94I, R204H and C266S.

To give but a few specific examples, when a modified mevalonate kinase polypeptide comprises 2 amino acid changes as compared with a parent mevalonate kinase polypeptide, it may comprise changes at positions corresponding to the following positions 132/375,167/169, 17/47 and/or 17/93 of SEQ ID NO: 1 of WO/2004/111214 (e.g. P132A/P375R, R167W/K169Q, I17T/G47D or I17T/K93E); when a modified mevalonate kinase polypeptide comprises 3 amino acid changes as compared with a parent mevalonate kinase, it may comprise changes at positions corresponding to the following positions 17/167/169, 17/132/375, 93/132/375, and/or 17/47/93 of SEQ ID NO: 1 of WO/2004/111214 (e.g., I17T/R167W/K169Q, I17T/P132A/P375R, K93E/P132A/P375R, I17T/R167W/K169H, I17T/R167T/K169M, I17T/R167T/K169Y, I17T/R167F/K169Q, I17T/R167I/K169N, I17T/R167H/K169Y, I17T/G47D/K93E or I17T/G47D/K93Q).

Thus, for example, a modified mevalonate kinase polypeptide may include one or more mutation(s) (particularly substitutions), as compared with a parent mevalonate kinase polypeptide, at one or more amino acid position (s) selected from the group consisting of amino acid positions corresponding to positions 55, 59, 66, 83, 106, 111, 117, 142, 152, 158, 218, 231, 249, 367 and 375 of the amino acid sequence of Saccharomyces cerevisiae mevalonate kinase as shown in SEQ ID NO: 1 of PCT application WO 2006/063752. For example, such corresponding substitutions may comprise one or more of P55L, F59S, N66K, C117S, or I152M. A modified mevalonate kinase may comprise a substitution corresponding to F59S substitution. A modified mevalonate kinase polypeptide comprising 2 amino acid changes as compared with its parent mevalonate kinase polypeptide may, for example, comprise changes at positions corresponding to the following positions 55/117, 66/152, 83/249, 111/375 or 106/218 of to SEQ ID NO: 1 of WO2006/063752 (e.g. P55L/C117S, N66K/1152M, K83E/S249P, H111N/K375N or L106P/S218P). A modified mevalonate kinase may comprise a substitution corresponding to N66K/I152M. A modified mevalonate kinase polypeptide comprising 4 amino acid changes as compared with its parent mevalonate kinase polypeptide may have changes at positions corresponding to one or more of the following positions 42/158/231/367 of SEQ ID NO:1 of WO2006/063752 (e.g. I142N/L158S/L231I/T367S).

According to the present invention, carotenoid production in a host organism may be adjusted by modifying the expression or activity of one or more proteins involved in isoprenoid biosynthesis. In some embodiments, such modification involves introduction of one or more heterologous isoprenoid biosynthesis polypeptides into the host cell; alternatively or additionally, modifications may be made to the expression or activity of one or more endogenous or heterologous isoprenoid biosynthesis polypeptides. Given the considerable conservation of components of the isoprenoid biosynthesis polypeptides, it is expected that heterologous isoprenoid biosynthesis polypeptides will often function even in significantly divergent organisms. Furthermore, should it be desirable to introduce more than one heterologous isoprenoid biosynthesis polypeptide (e.g., more than one version of the same polypeptide and/or more that one different polypeptides), in many cases polypeptides from different source organisms will function together. In some embodiments of the invention, a plurality of different heterologous isoprenoid biosynthesis polypeptides is introduced into the same host cell. In some embodiments, this plurality contains only polypeptides from the same source organism (e.g., two or more sequences of, or sequences derived from, the same source organism); in other embodiments the plurality includes polypeptides independently selected from from different source organisms (e.g., two or more sequences of, or sequences derived from, at least two independent source organisms).

In some embodiments of the present invention that utilize heterologous isoprenoid biosynthesis polypeptides, the source organisms include, but are not limited to, fungi of the genera Blakeslea, Candida, Cryptococcus, Cunninghamella, Lipomyces, Mortierella, Mucor, Phycomyces, Pythium, Rhodosporidium, Rhodotorula, Trichosporon, Yarrowia, Aspergillus, Botrytis, Cercospora, Fusarium (Gibberella), Kluyveromyces, Neurospora, Penicillium, Pichia (Hansenula), Puccinia, Saccharomyces, Schizosaccharomyces, Sclerotium, Trichoderms, Ustilago, and Xanthophyllomyces (Phaffia). In certain embodiments, the source organisms are of a species including, but not limited to, Cryptococcus neoformans, Fusarium fujikuroi, Kluyveromyces lactis, Neurospora crassa, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Ustilago maydis, and Yarrowia lipolytica.

As noted above, the isoprenoid biosynthesis pathway is also involved in the production of non-carotenoid compounds, such as sterols, steroids, and vitamins, such as vitamin E or vitamin K. Proteins that act on isoprenoid biosynthesis pathway intermediates, and divert them into biosynthesis of non-carotenoid compounds are therefore indirect inhibitors of carotenoid biosynthesis (see, for example, FIG. 5, which illustrates points at which isoprenoid intermediates are channeled into other biosynthesis pathways). Such proteins are therefore considered isoprenoid biosynthesis competitor polypeptides. Reductions of the level or activity of such isoprenoid biosynthesis competitor polypeptides are expected to increase carotenoid production in host cells according to the present invention. Additionally or alternatively, since disruption of the SAGA complex component SPT-8 increases carotenoid production (see e.g., Example 16), increased expression or activity of one or more components of the SAGA complex such as, without limitation, the SPT8 gene, may decrease production of carotenoids and/or retinolic compounds. Thus, polypeptides that comprise the SAGA complex can be considered competitor polypeptides in the situation where they decrease production of carotenoids and/or retinolic compounds. Without wishing to be bound by theory, the present invention encompasses the recognition that increased expression or activity of one or more components of the SAGA complex may act as isoprenoid biosynthesis competitors, thus reducing the amount of carotenoid produced. For example, one or more components of the SAGA complex may act on isoprenoid intermediates prior to GGPP, such that less GGPP is generated and available for the carotenoid generation pathway. In such embodiments, it will be understood that the SAGA polypeptide(s) components whose activity or expression is increased functions as isoprenoid biosynthesis competitor polypeptide(s). Thus, for example, one or more of the polypeptides encoded by the genes listed in Table 69 may function as isoprenoid biosynthesis competitor polypeptides. Such SAGA polypeptides can be expressed individually or in combination with one another. In certain embodiments, SAGA isoprenoid biosynthesis competitor polypeptides are expressed (and/or their activity increased) in combination with an increase in expression and/or activity of one or more additional isoprenoid biosynthesis competitor polypeptides, such as, without limitation, those isoprenoid biosynthesis competitor polypeptides listed in Tables 7-15.

In some embodiments of the present invention, production or activity of endogenous isoprenoid biosynthesis competitor polypeptides may be reduced or eliminated in host cells. In some embodiments, this reduction or elimination of the activity of an isoprenoid biosynthesis competitor polypeptide can be achieved by treatment of the host organism with small molecule inhibitors of enzymes of the ergosterol biosynthetic pathway. Enzymes of the ergosterol biosynthetic pathway include, for example, squalene synthase (Erg9), squalene epoxidase (Erg1), 2,3-oxidosqualene-lanosterol cyclase (Erg7), cytochrome P450 lanosterol 14α-demethylase (Erg11), C-14 sterol reductase (Erg24), C-4 sterol methyl oxidase (Erg25), SAM:C-24 sterol methyltransferase (Erg6), C-8 sterol isomerase (Erg2), C-5 sterol desaturase (Erg3), C-22 sterol desaturase (Erg5), and C-24 sterol reductase (Erg4) polypeptides. Each of these enzymes is considered an isoprenoid biosynthesis competitor polypeptide. Regulators of these enzymes may also be considered isoprenoid biosynthesis competitor polypeptides (e.g., the yeast proteins Sut1 (Genbank Accession JC4374 GI:2133159) and Mot3 (Genbank Accession NP_013786 GI:6323715), which may or may not have homologs in other organisms.

In other embodiments, reduction or elimination of the activity of an isoprenoid biosynthesis competitor polypeptide can be achieved by decreasing activity of the ubiquinone biosynthetic pathway. The commitment step in ubiquinone biosynthesis is the formation of para-hydroxybenzoate (PHB) from tyrosine or phenylalanine in mammals or chorismate in bacteria, followed by condensation of PHB and isoprene precursor, resulting in addition of the prenyl group. This reaction is catalyzed by PHB-polyprenyltransferase. The isoprenoid side chain of ubiquinone, which can be of varying length in different organisms, is determined by the prenyldiphosphate synthase enzyme. In organisms that produce the coenzyme Q10 form of ubiquinone, the 3-decaprenyl-4-hydroxybenzoic acid resulting from the condensation of PHB and decaprenyldiphosphate reaction undergoes further modifications, which include hydroxylation, methylation and decarboxylation, in order to form ubiquinone (CoQ10). Thus, reducing the activity of prenyldiphosphate synthase leading from farnesyldiphosphate to extended isoprenoids, or reducing the activity of PHB polyprenyltransferase may be useful in increasing the amount of isoprenoid available for carotenoid biosynthesis. (Examples of prenyldiphosphate synthase and PHB-polyprenyltransferase enzymes are depicted in Tables 29 and 30, respectively).

Known small molecule inhibitors of isoprenoid biosynthesis competitor enzymes include, but are not limited to, zaragosic acid (including analogs thereof such as TAN1607A (Biochem Biophys Res Commun 1996 Feb. 15; 219(2):515-520)), RPR 107393 (3-hydroxy-3-[4-(quinolin-6-yl)phenyl]-1-azabicyclo[2-2-2]octane dihydrochloride; J Pharmacol Exp Ther. 1997 May; 281(2):746-52), ER-28448 (5-{N-[2-butenyl-3-(2-methoxyphenyl)]-N-methylamino}-1,1-pentylidenebis(phosphonic acid) trisodium salt; Journal of Lipid Research, Vol. 41, 1136-1144, July 2000), BMS-188494 (The Journal of Clinical Pharmacology, 1998; 38:1116-1121), TAK-475 (1-[2-[(3R,5S)-1-(3-acetoxy-2,2-dimethylpropyl)-7-chloro-1,2,3,5-tetrahydro-2-oxo-5-(2,3-dimethoxyphenyl)-4,1-benzoxazepine-3-yl]acetyl]piperidin-4-acetic acid; Eur J Pharmacol. 2003 Apr. 11; 466(1-2):155-61), YM-53601 ((E)-2-[2-fluoro-2-(quinuclidin-3-ylidene)ethoxy]-9H-carbazole monohydrochloride; Br J Pharmacol. 2000 September; 131(1):63-70), or squalestatin I that inhibit squalene synthase; terbinafine (e.g., LAMISIL®), naftifine (NAFTIN®), S-allylcysteine, garlic, resveratrol, NB-598 (e.g., from Banyu Pharmaceutical Co), and/or green tea phenols that inhibit squalene epoxidase (see, for example, J. Biol Chem 265:18075, 1990; Biochem. Biophys. Res. Commun. 268:767, 2000); various azoles that inhibit cytochrome P450 lanosterol 14α-demethylase; and fenpropimorph that inhibits the C-14 sterol reductase and the C-8 sterol isomerase. In other embodiments, heterologous isoprenoid biosynthesis competitor polypeptides may be utilized (whether functional or non-functional; in some embodiments, dominant negative mutants are employed).

One particular isoprenoid biosynthesis competitor polypeptide useful according to the present invention is squalene synthase which has been identified and characterized from a variety of organisms; representative examples of squalene synthase polypeptide sequences are included in Table 16. In some embodiments of the invention that utilize squalene synthase (or modifications of squalene synthase) source organisms include, but are not limited to, Neurospora crassa, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), Aspergillus niger, Saccharomyces cerevisiae, Mucor circinelloides, Rhotorula glutinis, Candida utilis, Mortierella alpina, and Yarrowia lipolytica.

The carotenoid biosynthesis pathway branches off from the isoprenoid biosynthesis pathway at the point where GGPP is formed. The commitment step in carotenoid biosynthesis is the formation of phytoene by the head-to-head condensation of two molecules of GGPP, catalyzed by phytoene synthase (often called crtB; see FIG. 6). A series of dehydrogenation reactions, each of which increases the number of conjugated double bonds by two, converts phytoene into lycopene via neurosporene. The pathway branches at various points, both before and after lycopene production, so that a wide range of carotenoids can be generated. For example, action of a cyclase enzyme on lycopene generates γ-carotene; action of a desaturase instead produces 3,4-didehydrolycopene. γ-carotene is converted to β-carotene through the action of a cyclase. β-carotene can be processed into any of a number of products (see, for example, FIG. 6C), including astaxanthin (via echinenone, hydroxyechinenone, and phoenicoxanthin).

According to the present invention, carotenoid production in a host organism may be adjusted by modifying the expression or activity of one or more proteins involved in carotenoid biosynthesis. As indicated, in some embodiments, it will be desirable to utilize as host cells organisms that naturally produce one or more carotenoids. In some such cases, the focus will be on increasing production of a naturally-produced carotenoid, for example by increasing the level and/or activity of one or more proteins involved in the synthesis of that carotenoid and/or by decreasing the level or activity of one or more proteins involved in a competing biosynthetic pathway. Alternatively or additionally, in some embodiments it will be desirable to generate production of one or more carotenoids not naturally produced by the host cell.

According to some embodiments of the invention, it will be desirable to introduce one or more heterologous carotenogenic polypeptides into a host cell. As will be apparent to those of ordinary skill in the art, any of a variety of heterologous polypeptides may be employed; selection will consider, for instance, the particular carotenoid whose production is to be enhanced. The present invention contemplates not only introduction of heterologous carotenogenic polypeptides, but also adjustment of expression or activity levels of heterologous or endogenous carotenogenic polypeptides, including, for example, alteration of constitutive or inducible expression patterns. In some embodiments of the invention, expression patterns are adjusted such that growth in nutrient-limiting conditions is not required to induce oleaginy. For example, genetic modifications comprising alteration and/or addition of regulatory sequences (e.g., promoter elements, terminator elements) may be utilized to confer particular regulation of expression patterns. Such genetic modifications may be utilized in conjunction with endogenous genes (e.g., for regulation of endogenous carotenogenic); alternatively, such genetic modifications may be included so as to confer regulation of expression of at least one heterologous polypeptide (e.g., carotenogenic polypeptide(s)). For example, promoters including, but not limited to those described herein, such as Tef1, Gpd1 promoters can be used in conjunction with endogenous genes and/or heterolous genes for modification of expression patterns of endogenous carotenogenic polypeptide(s) and/or heterologous carotenogenic polypeptide(s). Similarly, exemplary terminator sequences include, but are not limited to, use of Y. lipolytica XPR2 terminator sequences.

As indicated in FIG. 6 and in the literature, proteins involved in carotenoid biosynthesis include, but are not limited to, phytoene synthase, phytoene dehydrogenase, lycopene cyclase, carotenoid ketolase, carotenoid hydroxylase, astaxanthin synthase (a single multifunctional enzyme found in some source organisms that typically has both ketolase and hydroxylase activities), carotenoid epsilon hydroxylase, lycopene cyclase (beta and epsilon subunits), carotenoid glucosyltransferase, and acyl CoA:diacyglycerol acyltransferase. Representative example sequences for these carotenoid biosynthesis polypeptides are provided in Tables 17a-25.

Alternatively or additionally, modified carotenoid ketolase polypeptides that exhibit improved carotenoid production activity may be utilized in accordance with the present invention. For example, carotenoid ketolase polypeptides comprising one more mutations to corresponding to those identified Sphingomonas sp. DC18 which exhibited improved astaxanthin production (Tao et al. 2006 Metab Eng. 2006 Jun. 27) and Paracoccus sp. strain N81106 which exhibited altered carotenoid production (Ye et al. 2006 Appl Environ Microbiol 72:5829-37).

Xanthophylls can be distinguished from other carotenoids by the presence of oxygen containing functional groups on their cyclic end groups. For instance, lutein and zeaxanthin contain a single hydroxyl group on each of their terminal ring structures, while astaxanthin contains both a keto group and a hydroxyl on each terminal ring. This property makes xanthophylls more polar than carotenes such as beta-carotene and lycopene, and thus dramatically reduces their solubility in fats and lipids. Naturally occurring xanthophylls are often found as esters of the terminal hydroxyl groups, both mono- and diesters of fatty acids. They also occur as glucosides in certain species of bacteria. The solubility and dispersibility of xanthophylls can be greatly modified by the addition of ester moieties, and it is known that esterification can also affect the absorbability and/or bioavailability of a given carotenoid. It is an objective of this invention to maximize the amount of a particular xanthophyll accumulating within the intracellular triacylglyceride fraction of oleaginous yeasts, and one mechanism for achieving this goal is to increase the hydrophobic nature of the xanthophyll product that accumulates. One way of achieving this is to engineer the production of fatty-acyl mono- and/or diesters of the target xanthophyll compound.

A variety of enzymes can function to esterify carotenoids. For example, carotenoid glucosyltransferases have been identified in several bacterial species (see, e.g., Table 24). In addition, acyl CoA:diacyglycerol acyltransferase (DGAT) and acyl CoA:monoacylglycerol acyltransferases (MGAT), which function in the final steps of triacylglycerol biosynthesis, are likely to serve an additional role in the esterification of xanthophylls. Representative DGAT polypeptides are shown in Table 25. Furthermore, other enzymes may specifically modify carotenoids and molecules of similar structure (e.g. sterols) and be available for modification and ester production.

In some embodiments of the invention, potential source organisms for carotenoid biosynthesis polypeptides include, but are not limited to, genera of naturally oleaginous or non-oleaginous fungi that naturally produce carotenoids. These include, but are not limited to, Botrytis, Cercospora, Fusarium (Gibberella), Mucor, Neurospora, Phycomyces, Puccina, Rhodotorula, Sclerotium, Trichoderma, and Xanthophyllomyces. Exemplary species include, but are not limited to, Neurospora crassa, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), Mucor circinelloides, and Rhodotorula glutinis. Of course, carotenoids are produced by a wide range of diverse organisms such as plants, algae, yeast, fungi, bacteria, cyanobacteria, etc. Any such organisms may be source organisms for carotenoid biosynthesis polypeptides according to the present invention.

In certain embodiments of the invention, carotenoid production in a host organism may be adjusted by modifying the activity of one or more endogenous genes that affect carotenoid biosynthesis. For example, as shown in Example 16, disruption of the endogenous SPT8 gene (YALI0E23804g) in Yarrowia lipolytica results in increased carotenoid production. SPT8 functions as part of the SAGA histone acetyltransferase complex, which is required for normal expression of some fungal genes and is thought to function as a coactivator complex in a multistep pathway leading to gene activation. Thus, without wishing to be bound by theory, the present invention encompasses the recognition that alteration of the expression and/or activity of one or more components of the SAGA histone acetyltransferase complex result in increased carotenoid production. Additionally, it will be appreciated by those of ordinary skill in the art that by increasing production of carotenoid(s) in a host organism by altering the expression and/or activity of one or more components of the SAGA histone acetyltransferase complex, production of a retinolic compound(s) in a host organism able to utilize such a carotenoid(s) as a substrate may also be increased since more of the cartenoid substrate will be available.

In Saccharomyces cerevisiae, the SAGA complex is a 1.8-MDa complex comprising a variety of components including distinct classes of transcription factors, such as Ada proteins (Ada1p, Ada2p, Ngg1p/Ada3p, and Ada4p/Gcn5p), TATA-binding protein (TBP)-related SPT proteins (Spt3p, Spt7p, Spt8p, and Spt20p/Ada5p), and TBP-associated factors or (TAFIIs) (TAFII90, TAFII68/61, TAFII60, TAFII25/23, and TAFII17). The SAGA complex also comprises the DNA-dependent protein kinase related molecule Tra1p, acetyltransferase and ubiquitin protease activities. The SAGA complex core comprises Ada and Spt subunits, a subset of Tafs, acetyltransferase and ubiquitin protease activities, the essential factor Tra1p, and two factors related to TBP function, Spt3 and Spt8. Several components of the Saccharomyces cerevisiae SAGA complex and their corresponding Yarrowia lipolytica homologs, are listed in Table 69. Each of these SAGA complex components is encompassed by the recombinant fungal strains, methods and compositions of the present invention. Those of ordinary skill in the art will be aware of these and other SAGA components, and will be able to modify such components in accordance with the present invention.

Certain SAGA components are essential. For example, in Saccharomyces cerevisiae, the TRA1 gene is essential. Thus, in certain embodiments, production of a carotenoid is increased by altering expression and/or activity of the TRA1 such that the host organism remains viable. For example, the expression and/or activity of the TRA1 gene or gene product may be decreased to a level below the expression and/or activity of wild type TRA1, but not to such an extent as to result in lethality. Those of ordinary skill in the art will be aware of tra1 mutations that result in decreased expression and/or activity but that do not result in lethality. Furthermore, it will be within the capability of one of ordinary skill in the art to identify such mutations without undue experimentation, for example by employing standard mutagenesis/screening techniques.

In certain embodiments of the present invention, production of one or more carotenoids is increased by alteration of the expression and/or activity of one or more components of the SAGA histone acetyltransferase complex in one or more of the following host organisms: Aspergillus, Blakeslea, Botrytis, Candida, Cercospora, Cryptococcus, Cunninghamella, Fusarium (Gibberella), Kluyveromyces, Lipomyces, Mortierella, Mucor, Neurospora, Penicillium, Phycomyces, Pichia (Hansenula), Puccinia, Pythium, Rhodosporidium, Rhodotorula, Saccharomyces, Sclerotium, Trichoderma, Trichosporon, Xanthophyllomyces (Phaffia), and Yarrowia; or is a species selected from the group consisting of: Aspergillus terreus, Aspergillus nidulans, Aspergillus niger, Blakeslea trispora, Botrytis cinerea, Candida japonica, Candida pulcherrima, Candida revkaufi, Candida tropicalis, Candida utilis, Cercospora nicotianae, Cryptococcus curvatus, Cunninghamella echinulata, Cunninghamella elegans, Fusarium fujikuroi (Gibberella zeae), Kluyveromyces lactis, Lipomyces starkeyi, Lipomyces lipoferus, Mortierella alpina, Mortierella ramanniana, Mortierella isabellina, Mortierella vinacea, Mucor circinelloides, Neurospora crassa, Phycomyces blakesleanus, Pichia pastoris, Puccinia distincta, Pythium irregulare, Rhodosporidium toruloides, Rhodotorula glutinis, Rhodotorula graminis, Rhodotorula mucilaginosa, Rhodotorula pinicola, Rhodotorula gracilis, Saccharomyces cerevisiae, Sclerotium rolfsii, Trichoderma reesei, Trichosporon cutaneum, Trichosporon pullans, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), and/or Yarrowia lipolytica.

In certain embodiments, production of one or more carotenoids is increased by altering expression and/or activity of one or more components of the SAGA histone acetyltransferase complex in a host organism, in combination with one or more additional carotenogenic modifications as described herein. For example, such one or more additional carotenogenic modifications may comprise heterologous expression of one or more carotenogenic polypeptides, isoprenoid biosynthesis polypeptides, carotenoid biosynthesis polypeptides, etc.

In certain embodiments, production of one or more carotenoids is increased by altering expression and/or activity of one or more components of the SAGA histone acetyltransferase complex in a host organism, in combination with one or more oleaginic modifications, as described herein. In certain embodiments, production of one or more carotenoids is increased by altering expression and/or activity of one or more components of the SAGA histone acetyltransferase complex in a host organism that is not naturally oleaginous. In certain embodiments, production of one or more carotenoids is increased by altering expression and/or activity of one or more components of the SAGA histone acetyltransferase complex in a host organism that is naturally oleaginous.

It will be appreciated that the particular carotenogenic modification to be applied to a host cell in accordance with the present invention will be influenced by which carotenoid(s) is desired to be produced. For example, isoprenoid biosynthesis polypeptides are relevant to the production of most carotenoids. Carotenoid biosynthesis polypeptides are also broadly relevant. Carotenoid ketolase activity is particularly relevant for production of canthaxanthin, as carotenoid hydroxylase activity is for production of lutein and zeaxanthin, among others. Both carotenoid hydroxylase and ketolase activities (and astaxanthin synthase) are particularly useful for production of astaxanthin.

In certain embodiments, host cells are engineered to produce carotenoids by introducing one or more carotenoid biosynthesis polypeptides. In general, any carotenoid biosynthesis polypeptide can be introduced into any host cell of the present invention. In certain embodiments, such carotenoid biosynthesis polypeptides are codon-optimized to accommodate the codon preferences of the host cell. In certain embodiments, a carotenoid biosynthesis polypeptide introduced into a host cell is from the same organism as the host cell and/or a related organism. For example, without limitation, the present invention encompasses the recognition that it may be desirable to introduce a fungal carotenoid biosynthesis polypeptide into a fungal host cell (e.g., from the same and/or a related fungal species). In certain embodiments, the host cell is a Y. lipolytica host cell. In certain aspects of such embodiments, a Y. lipolytica carotenoid biosynthesis polypeptide is introduced into the Y. lipolytica host cell. In certain aspects, a S. cerevisiae carotenoid biosynthesis polypeptide is introduced into the Y. lipolytica host cell. In certain aspects, any of a variety of fungal carotenoid biosynthesis polypeptides is introduced into the Y. lipolytica host cell.

Engineering Retinolic Compound Production

Retinolic compounds are synthesized from certain carotenoid precursors, which are themselves synthesized from isoprenoid precursors, some of which are also involved in the production of steroids and sterols (see description under section entitled “Engineering Carotenoid Production”). Thus, any carotenogenic modification that results in the increased production of a carotenoid from which a retinolic compound can be produced may similarly result in an increased production of a retinolic compound. Retinolic compounds comprise retinol, retinal, and retinoic acid, which together are collectively referred to as “Vitamin A”. In certain embodiments, the retinolic compound retinol is synthesized from the carotenoid precursor beta-carotene. Other carotenoid compounds that contain at least one beta-ionone ring structure, such as alpha-carotene and beta-cryptoxanthin, can be precursor compounds for synthesis of retinolic compounds.

According to the present invention, retinolic compound production in a host organism may be adjusted by modifying the expression or activity of one or more proteins involved in retinolic compound biosynthesis. As indicated, in some embodiments, it will be desirable to utilize as host cells organisms that naturally produce one or more retinolic compounds. In some such cases, the focus will be on increasing production of a naturally-produced retinolic compound, for example by increasing the level and/or activity of one or more proteins involved in the synthesis of that retinolic compound and/or by decreasing the level or activity of one or more proteins involved in a competing biosynthetic pathway. Alternatively or additionally, in some embodiments it will be desirable to generate production of one or more retinolic compounds not naturally produced by the host cell.

According to some embodiments of the invention, it will be desirable to introduce one or more heterologous retinologenic polypeptides into a host cell. As will be apparent to those of ordinary skill in the art, any of a variety of heterologous polypeptides may be employed; selection will consider, for instance, the particular retinolic compound whose production is to be enhanced. The present invention contemplates not only introduction of heterologous retinologenic polypeptides, but also adjustment of expression or activity levels of heterologous retinologenic polypeptides, including, for example, alteration of constitutive or inducible expression patterns. In some embodiments of the invention, expression patterns are adjusted such that growth in nutrient-limiting conditions is not required to induce oleaginy. For example, genetic modifications comprising alteration and/or addition of regulatory sequences (e.g., promoter elements, terminator elements) may be utilized to confer particular regulation of expression patterns. Such genetic modifications may be utilized in conjunction with endogenous genes (e.g., for regulation of endogenous carotenogenic); alternatively, such genetic modifications may be included so as to confer regulation of expression of at least one heterologous polypeptide (e.g., retinologenic polypeptide(s)). For example, promoters including, but not limited to those described herein, such as Tef1, Gpd1 promoters can be used in conjunction with endogenous genes and/or heterolous genes for modification of expression patterns of endogenous retinologenic polypeptide(s) and/or heterologous retinologenic polypeptide(s). Similarly, exemplary terminator sequences include, but are not limited to, use of Y. lipolytica XPR2 terminator sequences.

As indicated in FIG. 11 and in the literature, proteins involved in retinologenic biosynthesis include, but are not limited to, beta-carotene 15,15′-monooxygenase (also known as beta-carotene dioxygenase) and beta carotene retinol dehydrogenase. Representative example sequences for these retinolic compound biosynthesis polypeptides are provided in Tables 67-68.

In some embodiments of the invention, potential source organisms for retinolic compound biosynthesis polypeptides include, but are not limited to, genera of naturally oleaginous or non-oleaginous fungi that naturally produce carotenoids. These include, but are not limited to, Botrytis, Cercospora, Fusarium (Gibberella), Mucor, Neurospora, Phycomyces, Puccina, Rhodotorula, Sclerotium, Trichoderma, and Xanthophyllomyces. Exemplary species include, but are not limited to, Neurospora crassa, Xanthophyllomyces dendrorhous (Phaffia rhodozyma), Mucor circinelloides, and Rhodotorula glutinis. Of course, retinolic compounds are produced by a wide range of diverse organisms such as mammals, bacteria, cyanobacteria, etc. Any such organisms may be source organisms for retinolic compound biosynthesis polypeptides according to the present invention.

In certain embodiments of the invention, retinolic compound production in a host organism that is able to produce retinolic compounds from carotenoid substrates is adjusted by modifying the activity of one or more endogenous genes that affect carotenoid biosynthesis. For example, as shown in Example 16, disruption of the endogenous SPT8 gene (YALI0E23804g) in Yarrowia lipolytica results in increased carotenoid production. As will be appreciated by those of ordinary skill in the art, increasing production of a carotenoid(s) in a host organism by altering the expression and/or activity of one or more components of the SAGA histone acetyltransferase complex will result in a greater abundance of such a carotenoid(s); hence, production of a retinolic compound(s) in a host organism able to utilize such a carotenoid(s) as a substrate may similarly be increased.

Without wishing to be bound by theory, the present invention contemplates that alteration of the expression and/or activity of one or more components of the SAGA histone acetyltransferase complex may result in increased retinolic compound production. In certain embodiments, retinolic compound production is increased in a host organism by altering the expression and/or activity of one or more of: Ada proteins (Ada1p, Ada2p, Ngg1p/Ada3p, and Ada4p/Gcn5p), TATA-binding protein (TBP)-related SPT proteins (Spt3p, Spt7p, Spt8p, and Spt20p/Ada5p), TBP-associated factors or (TAFIIs) (TAFII90, TAFII68/61, TAFII60, TAFII25/23, and TAFII17), Tra1p, and/or proteins encoding the acetyltransferase and/or ubiquitin protease activities. In certain embodiments, retinolic compound production is increased in a host organism by altering the expression and/or activity of one or more polypeptides listed in Table 69. Those of ordinary skill in the art will be aware of these and other SAGA components, and will be able to modify such components in accordance with the present invention.

In certain embodiments, host cells are engineered to produce retinolic compounds by introducing one or more carotenoid biosynthesis polypeptides. In general, any retinolic compound biosynthesis polypeptide can be introduced into any host cell of the present invention. In certain embodiments, such retinolic compound biosynthesis polypeptides are codon-optimized to accommodate the codon preferences of the host cell. In certain embodiments, a retinolic compound biosynthesis polypeptide introduced into a host cell is from the same organism as the host cell and/or a related organism. For example, without limitation, the present invention encompasses the recognition that it may be desirable to introduce a fungal retinolic compound biosynthesis polypeptide into a fungal host cell (e.g., from the same and/or a related fungal species). In certain embodiments, the host cell is a Y. lipolytica host cell. In certain aspects of such embodiments, a Y. lipolytica retinolic compound biosynthesis polypeptide is introduced into the Y. lipolytica host cell. In certain aspects, a S. cerevisiae retinolic compound biosynthesis polypeptide is introduced into the Y. lipolytica host cell. In certain aspects, any of a variety of fungal retinolic compound biosynthesis polypeptides is introduced into the Y. lipolytica host cell.

Production and Isolation of Carotenoids and/or Retinolic Compounds

As discussed above, accumulation of lipid bodies in oleaginous organisms is generally induced by growing the relevant organism in the presence of excess carbon source and limiting nitrogen and/or other nutrients (eg. phosphate and magnesium). Specific conditions for inducing such accumulation have previously been established for a number of different oleaginous organisms (see, for example, Wolf (ed.) Nonconventional yeasts in biotechnology Vol. 1, Springer-Verlag, Berlin, Germany, pp. 313-338; Lipids 18(9):623, 1983; Indian J. Exp. Biol. 35(3):313, 1997; J. Ind. Microbiol. Biotechnol. 30(1):75, 2003; Bioresour Technol. 95(3):287, 2004, each of which is incorporated herein by reference in its entirety).

In general, it will be desirable to cultivate inventive modified host cells under conditions that allow accumulation of at least about 20% of their dry cell weight as lipid. In other embodiments, the inventive modified host cells are grown under conditions that permit accumulation of at least about 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or even 80% or more of their dry cell weight as lipid. In certain embodiments, the host cells utilized are cells which are naturally oleaginous, and induced to produce lipid to the desired levels. In other embodiments, the host cells are cells which naturally produce lipid, but have been engineered to increase production of lipid such that desired levels of lipid production and accumulation are achieved.

In certain embodiments, the host cells of the invention are not naturally oleaginous, but have been engineered to produce lipid such that desired levels of lipid production are obtained. Those of ordinary skill in the art will appreciate that, in general, growth conditions that are effective for inducing lipid accumulation in a source organism, may well also be useful for inducing lipid accumulation in a host cell into which the source organism's oleaginic polypeptides have been introduced. Of course, modifications may be required in light of characteristics of the host cell, which modifications are within the skill of those of ordinary skill in the art.

It will also be appreciated by those of ordinary skill in the art that it will often be desirable to ensure that production of the desired carotenoid and/or retinolic compound by the inventive modified host cell occurs at an appropriate time in relation to the induction of oleaginy such that the carotenoid(s) and/or retinolic compound(s) accumulate(s) in the lipid bodies. In some embodiments, it will be desirable to induce production of the carotenoid(s) and/or retinolic compound(s) in a host cell which does not naturally produce the carotenoid(s) and/or retinolic compound(s), such that detectable levels of the carotenoid(s) and/or retinolic compound(s) is/are produced. In certain aspects the host cells which do not naturally produce a certain carotenoid(s) and/or retinolic compound(s) are capable of production of other carotenoid(s) (e.g. certain host cells may, for example, naturally produce β-carotene but may not naturally produce astaxanthin) and/or retinolic compound(s), (e.g. certain host cells may, for example, naturally produce retinal but may not naturally produce retinol); in other aspects the host cells do not naturally produce any carotenoid(s) and/or retinolic compound(s). In other embodiments, it will be desirable to increase production levels of carotenoid(s) and/or retinolic compound(s) in a host cell which does naturally produce low levels of the carotenoid(s) and/or retinolic compound(s), such that increased detectable levels of the carotenoid(s) and/or retinolic compound(s) are produced. In certain aspects, the host cells which do naturally produce the carotenoid(s) (e.g., β-carotene) also produce additional carotenoid(s) (e.g., astaxanthin, etc.) and/or retinolic compound(s) (e.g., retinal); in still other aspects, the cells which naturally produce the carotenoid(s) (e.g., β-carotene) do not produce additional carotenoid(s) and/or retinolic compound(s).

In certain embodiments of the invention, it will be desirable to accumulate carotenoids and/or retinolic compounds to levels (i.e., considering the total amount of all produced carotenoids and/or retinolic compounds together or considering a particular carotenoid and/or retinolic compound) that are greater than at least about 1% of the dry weight of the cells. In some embodiments, the total carotenoid and/or retinolic compound accumulation will be to a level at least about 1.5%, at least about 2%, at least about 2.5%, at least about 3%, at least about 3.5%, at least about 4%, at least about 4.5%, at least about 5%, at least about 5.5%, at least about 6%, at least about 6.5%, at least about 7%, at least about 7.5%, at least about 8%, at least about 8.5%, at least about 9%, at least about 9.5%, at least about 10%, at least about 10.5%, at least about 11%, at least about 11.5%, at least about 12%, at least about 12.5%, at least about 13%, at least about 13.5%, at least about 14%, at least about 14.5%, at least about 15%, at least about 15.5%, at least about 16%, at least about 16.5%, at least about 17%, at least about 17.5%, at least about 18%, at least about 18.5%, at least about 19%, at least about 19.5%, at least about 20% or more of the total dry weight of the cells.

In some embodiments of the invention, a particular carotenoid and/or retinolic compound may not accumulate to a level as high as 1% of the total dry weight of the cells; appropriately engineered cells according to the present invention, and any lipid bodies, carotenoids and/or retinolic compounds they produce, remain within the scope of the present invention. Thus, in some embodiments, the cells accumulate a given carotenoid and/or retinolic compound to a level below about 1% of the dry weight of the cells. In some embodiments, the carotenoid and/or retinolic compound accumulates to a level below about 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.05%, or lower, of the dry cell weight of the cells.

In some embodiments of the invention, carotenoids and/or retinolic compounds accumulate both within lipid bodies and elsewhere in the cells. In some embodiments, carotenoids and/or retinolic compounds accumulate primarily within lipid bodies. In some embodiments, carotenoids and/or retinolic compounds accumulate substantially exclusively within lipid bodies. In some embodiments, at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more of a desired produced carotenoid and/or retinolic compound accumulates in lipid bodies.

In some embodiments of the invention, modified host cells are engineered to produce one or more carotenoid compound(s) and/or retinolic compound(s) characterized by negligible solubility in water (whether hot or cold) and detectable solubility in one or more oils. In some embodiments, such compounds have a solubility in oil below about 0.2%. In some embodiments, such compounds have a solubility in oil within the range of about <0.001%-0.2%.

The present invention therefore provides engineered host cells (and methods of making and using them) that contain lipid bodies and that further contain one or more carotenoid compounds and/or retinolic compounds accumulated in the lipid bodies, where the compounds are characterized by a negligible solubility in water and a solubility in oil within the range of about <0.001%-0.2%; 0.004%-0.15%; 0.005-0.1%; or 0.005-0.5%. For example, in some embodiments, such compounds have a solubility in oil below about 0.15%, 0.14%, 0.13%, 0.12%, 0.11%, 0.10%. 0.09, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.05%, or less. In some embodiments, the compounds show such solubility in an oil selected from the group consisting of sesame; soybean; apricot kernel; palm; peanut; safflower; coconut; olive; cocoa butter; palm kernel; shea butter; sunflower; almond; avocado; borage; carnauba; hazel nut; castor; cotton seed; evening primrose; orange roughy; rapeseed; rice bran; walnut; wheat germ; peach kernel; babassu; mango seed; black current seed; jojoba; macademia nut; sea buckthorn; sasquana; tsubaki; mallow; meadowfoam seed; coffee; emu; mink; grape seed; thistle; tea tree; pumpkin seed; kukui nut; and mixtures thereof.

Bacterial carotenogenic genes have already been demonstrated to be transferable to other organisms, and are therefore particularly useful in accordance with the present invention (see, for example, Miura et al., Appl. Environ. Microbiol. 64:1226, 1998). In other embodiments, it may be desirable to utilize genes from other source organisms such as plant, alga, or microalgae; these organisms provide a variety of potential sources for ketolase and hydroxylase polypeptides. Still additional useful source organisms include fungal, yeast, insect, protozoal, and mammalian sources of polypeptides.

In some embodiments of the present invention, isoprenoid production is increased in host cells (e.g., in Y. lipolytica cells) through expression of a truncated variant of a hydroxymethylglutaryl-CoA (HMG CoA) reductase polypeptide. In some embodiments, the truncated variant is a truncated variant of a Y. lipolytica HMG CoA reductase polypeptide. According to the present invention, expression of such a truncated HMG CoA reductase polypeptide can result in increased isoprenoid and/or carotenoid production in host cells (e.g., Y. lipolytica cells).

Alternatively or additionally, in some embodiments of the present invention, isoprenoid production is increased in host cells (e.g., in Y. lipolytica cells) through application of one or more carotenogenic modification(s) that increase(s) level and/or activity of a polypeptide selected from the group consisting of farnesyl pyrophosphate synthase polypeptides, geranylgeranylpyrophosphate synthase polypeptides, and combinations thereof. In some embodiments, the source organism for the selected polypeptide is Y. lipolytica.

Alternatively or additionally, in some embodiments of the present invention, isoprenoid production is increased in host cells (e.g., in Y. lipolytica cells) through application of one or more carotenogenic modification(s) that decrease(s) expression or activity of an isoprenoid biosynthesis competitor polypeptide (e.g., of a squalene synthase polypeptide), for example thereby reducing diversion of one or more intermediates away from the isoprenoid and/or carotenoid biosynthesis pathways. In some embodiments, the polypeptide whose expression or activity is reduced is endogenous to the host cell.

In some embodiments of the present invention, more than one carotenogenic modification is applied to the same host cell. For example, isoprenoid production may be increased in host cells (e.g., Y. lipolytica cells) through application of at least two or more carotenogenic modifications selected from the group consisting of: expression of a truncated HMG CoA reductase polypeptide, increase in expression and/or activity of farnesyl pyrophosphate synthase polypeptide, increase in expression and/or activity of a geranylgeranylpyrophosphate synthase polypeptide, decrease in expression and/or activity of a squalene synthase polypeptide, and combinations thereof.

Furthermore, in some embodiments of the invention, carotenoid production (e.g., production of β-carotene) is increased in host cells (e.g., in Y. lipolytica cells) through application of one or more carotenogenic modification(s) that increase(s) expression and/or activity of a polypeptide selected from the group consisting of phytoene synthase, lycopene cyclase, phytoene dehydrogenase, and combinations thereof. In some embodiments, such increase in expression comprises introduction of one or more genes encoding heterologous polypeptides. In some embodiments, phytoene synthase and lycopene cyclase activities are provided in a single polypeptide or complex (e.g., by the Mucor circinelloides or Neurospora crassa multifunctional phytoene synthase/lycopene cyclase). In some embodiments, phytoene dehydrogenase from Mucor circinelloides or Neurospora crassa is utilized.

In some embodiments, production of one or more carotenoids downstream of β-carotene (e.g., of one or more hydroxylated xanthophylls) is increased in host cells that produce β-carotene (including host cells that have been engineered to produce β-carotene, e.g., through application of one or more carotenogenic modifications as described herein) through application of one or more carotenogenic modifications that increase(s) level and/or activity of one or more carotenoid ketolase polypeptides (e.g., from Parvularcula bermudensis and/or Aurantimonase sp. SI85-9A1) to produce one or more ketone-containing carotenoids (e.g., canthaxanthin, echinenone, astaxanthin, and combinations thereof).

In some embodiments, production of one or more hydroxylated carotenoids is increased in host cells that produce (including having been engineered to produce) β-carotene and/or one or more ketone-containing carotenoids though application of one or more carotenogenic modifications that increase(s) the level and/or activity of one or more carotenoid hydroxylase polypeptides (e.g., from Xanthobacter autotrophicus and/or Erythrobacter litoralis) to increase production of one or more hydroxylated carotenoids (e.g., zeaxanthin, lutein, β-cryptoxanthin, astaxanthin, and combinations thereof).

Similar approaches to enhance carotenoid production may be employed in other oleaginous or non-oleaginous host organisms (e.g. S. cerevisiae, C. utilis, P. rhodozyma) can be undertaken, using the same, homologous, or functionally similar carotogenic polypeptides.

In some embodiments, the present invention provides modified Y. lipolytica strains that have been engineered to express one or more carotenoid biosynthesis polypeptides and/or isoprenoid biosynthesis polypeptides. For example, in some embodiments, a modified Y. lipolytica strain is engineered to increase expression and/or activity of one or more of phytoene synthase, phytoene dehydrogenase, lycopene cyclase, and GGPP synthase, and/or to decrease expression and/or activity of squalene synthase. In some embodiments, a modified Y. lipolytica strain is engineered to express all of these polypeptides. Such a modified Y. lipolytica strain produces β-carotene (see, for example, Example 2).

In some embodiments, inventive modified Y. lipolytica strains that have been engineered to produce β-carotene are further engineered to express a truncated HMG CoA reductase; in some such embodiments, the strains are engineered so that expression of the truncated HMG CoA reductase increases β-carotene several fold (for example, 3-4 fold or more).

In some embodiments, inventive modified Y. lipolytica strains that have been engineered to produce β-carotene are further engineered to express a beta-carotene 15,15′-monooxygenase and/or a retinol dehydrogenase to increase retinolic compound production.

In some embodiments, inventive modified Y. lipolytica strains that have been engineered to produce β-carotene are further engineered to express carotenoid hydroxylase (to achieve production of zeaxanthin and/or β-cryptoxanthin), carotenoid ketolase (to achieve production of canthaxanthin and/or echinenone), or both (to achieve production of astaxanthin).

In some embodiments, inventive modified Y. lipolytica strains that have been engineered to produce, for example, β-carotene, zeaxanthin, canthaxanthin, echinenone, and/or astaxanthin are also engineered to have increased expression of, for example, malic enzyme, mevalonate kinase, etc.

It will be appreciated that, in some embodiments of the invention, it may be desirable to engineer a particular host cell by expressing more than one version of a given polypeptide (e.g., isoprenoid biosynthesis polypeptide, carotenoid biosynthesis polypeptide, oleaginic polypeptide, isoprenoid biosynthesis competitor polypeptides, retinolic compound biosynthesis polypeptide, etc.). For example, a given host cell may be engineered to express versions of a given polypeptide from two or more different sources. Where a particular enzyme may be comprised of more than one polypeptide chains, it will often be desirable to utilize all chains from a single source, although this is not required so long as activity is achieved. Also, whenever a host cell is engineered to express a polypeptide from a different source, it may be desirable to alter the gene sequence encoding the polypeptide to account for codon preferences of the host cell.

To give but a few specific examples, the present invention provides modified Y. lipolytica strains that have been engineered to express the phytoene synthase/lycopene cyclase bifunctional (carB) polypeptide from M. circinelloides (see, for example, Example 1B), and also to express the phytoene dehydrogenase (carRP) polypeptide from M. circinelloides (see, for example, Example 1A). In some embodiments, the present invention provides such carB+carRP-expressing Y. lipolytica strains that have been engineered to modify expression and/or activity of a truncated HMG-CoA reductase polypeptide from Y. lipolytica and/or one or more Y. lipolytica polypeptides selected from the group consisting of GGPP synthase, FPP synthase (Erg20), IPP isomerase (IDI), HMG synthase (Erg13), mevalonate kinase (Erg12), squalene synthase (Erg9), phosphimevalonate kinase (Erg8), mevalonate pyrophosphate decarboxylase (MVD1), malic enzyme, malate dehydrogenase, glucose 6 phosphate dehydrogenase, malate dehydrogenase homolog 2,6-phosphogluconate dehydrogenase (GND1), isocitrate dehydrogenase, fructose 1,6 bisphosphatase, acetoacetyl CoA thiolase (Erg10), ATP citrate lyase subunit 1, ATP citrate lyase subunit 2, and combinations thereof. The present invention therefore specifically provides Y. lipolytica strains that have been engineered to produce β-carotene.

The present invention also specifically provides modified Y. lipolytica strains that have been engineered to express at least one carotenoid ketolase (e.g., crtO/crtW) polypeptide, and in some embodiments more than one, for example from a source selected from the group consisting of Parvularcula bermudensis (see, for example, Example 1H), Aurantimonas (see, for example, Example 1G), and/or an environmental isolate identified from the Sargasso Sea (see, for example, Example 1F). The present invention therefore specifically provides Y. lipolytica strains that have been engineered to produce canthaxanthin, astaxanthin, and/or echinenone.

The present invention further specifically provides modified Y. lipolytica strains that have been engineered to express at least one carotenoid hydroxylase (e.g., crtZ) polypeptide, and in some embodiments more than one, from Erythrobacter litoralis (see, for example, Examples 1J and 1L), Novosphingobium aromaticivarans (see, for example, Example 1E), Parvularcula bermudensis (see, for example, Example 1I), Xanthobacter autotrophicus (see, for example, Example 1O), Sphingopyxis alaskensis (see, for example, Example 1M), Chlamydomonas rheinhardtii, Erythrobacter longus, Robiginitalea biformata (see, or example, Example 1N) and/or Pseudomonas putida (see, for example, Example 1P). The present invention therefore specifically provides Y. lipolytica strains that have been engineered to produce zeaxanthin, lutein, β-cryptoxanthin, and/or astaxanthin.

The present invention further specifically provides modified Y. lipolytica strains that have been engineered to express at least one carotenoid ketolase (e.g., crtO/crtW) polypeptide in combination with at least one carotenoid hydroxylase (e.g., crtZ) polypeptide. In certain embodiments, the at least one carotenoid ketolatse polypeptide and at least one carotenoid hydroxylase polypeptide are encoded by nucleic acid sequences present in separate nucleic acid molecules. In certain embodiments, the at least one carotenoid ketolatse polypeptide and at least one carotenoid hydroxylase polypeptide are encoded by nucleic acid sequences present in the same nucleic acid molecule. For example, a host organism may be transformed or transfected with a single expression vector, which expression vector comprises both a carotenoid ketolatse polypeptide and a carotenoid hydroxylase polypeptide, each of which comprises sequences sufficient to direct their expression in the host organism.

In certain embodiments, the at least one carotenoid ketolase (e.g., crtO/crtW) polypeptide and the at least one carotenoid hydroxylase (e.g., crtZ) polypeptide are expressed as a fusion protein. A representative example of such embodiments is presented in Example 17. In certain embodiments, such a fusion polypeptide is designed such that the carotenoid ketolatse polypeptide is positioned N-terminal to the carotenoid hydroxylase polypeptide. In certain embodiments, such a fusion polypeptide is designed such that the carotenoid ketolatse polypeptide is positioned C-terminal to the carotenoid hydroxylase polypeptide.

In embodiments in which the carotenoid ketolatse polypeptide and the carotenoid hydroxylase polypeptide are expressed concurrently (whether from separate nucleic acid molecules or from the same nucleic acid molecule), the polypeptides may be selected from any of a variety of source organisms. As non-limiting examples, the carotenoid hydroxylase polypeptide may be selected from an organism such as Erythrobacter litoralis (see, for example, Examples 1J and 1L), Novosphingobium aromaticivarans (see, for example, Example 1E), Parvularcula bermudensis (see, for example, Example 1I), Xanthobacter autotrophicus (see, for example, Example 1O), Sphingopyxis alaskensis (see, for example, Example 1M), Chlamydomonas rheinhardtii, Erythrobacter longus, Robiginitalea biformata (see, or example, Example 1N) and/or Pseudomonas putida (see, for example, Example 1P). As further non-limiting examples, the carotenoid ketolase polypeptide may be selected from an organism such as Parvularcula bermudensis (see, for example, Example 1H), Aurantimonas (see, for example, Example 1G), and/or an environmental isolate identified from the Sargasso Sea (see, for example, Example 1F).

It should be noted that, for inventive organisms that produce more than one carotenoid, it will sometimes be possible to adjust the relative amounts of individual carotenoids produced by adjusting growth conditions. For example, it has been reported that controlling the concentration of dissolved oxygen in a culture during cultivation can regulate relative production levels of certain carotenoids such as β-carotene, echinenone, β-cryptoxanthin, 3-hydroxyechinenone, asteroidenone, canthaxanthin, zeaxanthin, adonirubin, adonixanthin and astaxanthin (see, for example, U.S. Pat. No. 6,825,002 to Tsubokura et al., the entire contents of which are incorporated herein by reference). Additionally or alternatively, the present invention encompasses the recognition that controlling the pH in a culture during cultivation can regulate relative production levels of these and/or other carotenoids (see e.g., Example 18).

Particularly for embodiments of the present invention directed toward production of astaxanthin, it will often be desirable to utilize one or more genes from a natural astaxanthin-producing organism. Where multiple heterologous polypeptides are to be expressed, it may be desirable to utilize the same source organism for all, or to utilize closely related source organisms.

Inventive modified cells, that have been engineered to produce carotenoids and/or to accumulate lipid (including to be oleaginous), can be cultured under conditions that achieve carotenoid production and/or oleaginy. In some embodiments, it will be desirable to control growth conditions so in order to maximize production of a particular carotenoid or set of carotenoids (including all carotenoids) and/or to optimize accumulation of the particular carotenoid(s) in lipid bodies. In some embodiments it will be desirable to control growth conditions to adjust the relative amounts of different carotenoid products produced.

Inventive modified cells, that have been engineered to produce retinolic compounds and/or to accumulate lipid (including to be oleaginous), can be cultured under conditions that achieve retinolic compound production and/or oleaginy. In some embodiments, it will be desirable to control growth conditions so in order to maximize production of a particular retinolic compound or set of retinolic compounds (including all retinolic compounds) and/or to optimize accumulation of the particular retinolic compound (s) in lipid bodies. In some embodiments it will be desirable to control growth conditions to adjust the relative amounts of different retinolic compound products produced.

In some embodiments, it will be desirable to limit accumulation of a particular intermediate, for example ensuring that substantially all of a particular intermediate compound is converted so that accumulation is limited. For example, particularly in situations where a downstream enzyme may be less efficient than an upstream enzyme and it is desirable to limit accumulation of the product of the upstream enzyme (e.g., to avoid its being metabolized via a competitive pathway and/or converted into an undesirable product), it may be desirable to grow cells under conditions that control (e.g., slow) activity of the upstream enzyme so that the downstream enzyme can keep pace.

Those of ordinary skill in the art will appreciate that any of a variety of growth parameters, including for example amount of a particular nutrient, pH, temperature, pressure, oxygen concentration, timing of feeds, content of feeds, etc can be adjusted as is known in the art to control growth conditions as desired.

To give but a few examples, in some embodiments, growth and/or metabolism is/are limited by limiting the amount of biomass accumulation. For example, growth and/or metabolism can be limited by growing cells under conditions that are limiting for a selected nutrient. The selected limiting nutrient can then be added in a regulated fashion, as desired. In some embodiments, the limiting nutrient is carbon, nitrogen (e.g., via limiting ammonium or protein), phosphate, magnesium, one or more trace metals, or combinations thereof. In some embodiments, the limiting nutrient is carbon. In some embodiments, the limiting nutrient is one or more trace metals.

In some embodiments, use of a limiting nutrient can by utilized to control metabolism of a particular intermediate and/or to adjust relative production of particular carotenoid compounds and/or retinolic compounds. In some embodiments, this result can be achieved by controlling metabolism of a particular intermediate as discussed above; in some embodiments, it can be achieved, for example, by limiting progress through the carotenoid and/or retinolic compound biosynthesis pathway so that a desired carotenoid product (e.g., β-carotene, canthaxanthin, astaxanthin, etc.) or retinolic compound (e.g., retinal) is not converted to a downstream compound. To give but one example, phosphate limitation can slow the overall rate of clux through the carotenoid biosynthesis pathway and can be utilized to change the ratio of canthaxanthin to echinenone produced.

In some embodiments, cells are grown in the presence of excess carbon source and limiting nitrogen, phosphate, and/or magnesium to induce oleaginy. In some embodiments cells are grown in the presence of excess carbon source and limiting nitrogen. In some embodiments, the carbon:nitrogen ratio is within the range of about 200:1, 150:1, 125:1, 100:1, 95:1, 90:1, 85:1, 80:1, 75:1, 70:1, 65:1, 60:1, 55:1, 50:1, 45:1, 40:1, 35:1, 30:1, 25:1, 20:1, 15:1, 10:1, or less. Those of ordinary skill in the art are aware of a wide variety of carbon sources, including, for example, glycerol, glucose, galactose, dextrose, any of a variety of oils (e.g., olive, canola, corn, sunflower, soybean, cottonseed, rapeseed, etc., and combinations thereof) that may be utilized in accordance with the present invention. Combinations of such may also be utilized. For example, common carbon source compositions contain oil:glucose in a ratio within the range of about 5:95 to 50:50 (e.g. about 5:95, about 10:90, about 15:85, about 20:80, about 25:75, about 30:70, about 35:65, about 40:60, about 45:55, about 50:50).

Those of ordinary skill in the art are also aware of a variety of different nitrogen sources (e.g., ammonium sulfate, proline, sodium glutamate, soy acid hydrolysate, yeast extract-peptone, yeast nitrogen base, corn steep liquor, etc, and combinations thereof) that can be utilized in accordance with the present invention.

In some embodiments, cultures are grown at a selected oxygen concentration (e.g., within a selected range of oxygen concentrations). In some embodiments, oxygen concentration may be varied during culture. In some embodiments, oxygen concentration may be controlled during some periods of culture and not controlled, or controlled at a different point, during others. In some embodiments, oxygen concentration is not controlled. In some embodiments, cultures are grown at an oxygen concentration within the range of about 5-30%, 5-20%, 10-25%, 10-30%, 15-25%, 15-30%, including at about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or more. In some embodiments, oxygen concentration is maintained above about 20%, at least for some period of the culture.

In some embodiments, cells are grown via fed-batch fermentation. In some embodiments, feed is continued until feed exhaustion and/or the feed is controlled to initiate or increase once a certain level of dissolved oxygen is detected in the culture medium (e.g., about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, or more dissolved oxygen). The feed rate can be modulated to maintain the dissolved oxygen at a specific level (e.g., about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, or more dissolved oxygen).

In some embodiments, inventive modified cells are grown in a two-phase feeding protocol in which the first phase is designed to maintain conditions of excess carbon and limiting oxygen, and the second phase results in conditions of excess oxygen and limiting carbon. The carbon sources in each phase can be the same (e.g., both glucose) or different (e.g., glucose then glucose-oil mixture, oil then glucose, or glucose-oil mixture then glucose). The present invention demonstrates that such conditions can achieve high levels of carotenoid production (see, for example, Example 5D). Additionally or alternatively, such conditions also result in high levels of retinolic compound production. For example, high levels of retinolic compound(s) production may be achieved by increasing the levels of a particular carotenoid that is used as a substrate for the production of such a retinolic compound(s).

In some embodiments, inventive modified cells are cultivated at constant temperature (e.g., between about 20-40, or 20-30 degrees, including for example at about 20, 20.5, 21, 21.5, 22, 22.5, 23, 23.5, 24, 24.5, 25, 25.5, 26, 26.5, 27, 27.5, 28, 28.5, 29, 29.5, 30° C. or above) and/or pH (e.g., within a range of about 4-7.5, or 4-6.5, 3.5-7, 3.5-4, 4-4.5, 4.5-5, 5-5.5, 5.5-6, 6-6.5, 6.5-7, 7-7.5, 7-8, etc., including at about 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5 or above); in other embodiments, temperature and/or pH may be varied during the culture period, either gradually or in a stepwise fashion.

For example, in some embodiments, the pH is 7.0 at inoculation and is increased to pH 8.0 during the course of the fermentation. The pH may be increased either continuously or in discrete steps. For example, in Example 19, the pH of the culture in increased continuously. In certain embodiments, the pH in increased continuously by increasing the pH at a rate of 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.010, 0.011, 0.012, 0.013, 0.014, 0.015, 0.016, 0.017, 0.018, 0.019, 0.020, 0.021, 0.022, 0.023, 0.024, 0.025, 0.026, 0.027, 0.028, 0.029, 0.030, 0.031, 0.032, 0.033, 0.034, 0.035, 0.036, 0.037, 0.038, 0.039, 0.040, 0.041, 0.042, 0.043, 0.044, 0.045, 0.046, 0.047, 0.048, 0.049, 0.050 or more units/hour.

In certain embodiments, the pH in increased in discrete steps by increasing the pH by 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.010, 0.011, 0.012, 0.013, 0.014, 0.015, 0.016, 0.017, 0.018, 0.019, 0.020, 0.021, 0.022, 0.023, 0.024, 0.025, 0.026, 0.027, 0.028, 0.029, 0.030, 0.031, 0.032, 0.033, 0.034, 0.035, 0.036, 0.037, 0.038, 0.039, 0.040, 0.041, 0.042, 0.043, 0.044, 0.045, 0.046, 0.047, 0.048, 0.049, 0.050 or more at each step.

In certain embodiments, the pH is increased employing a combination of continuous increase and discrete steps.

In certain embodiments, increasing the pH during the course of fermentation results in one or more beneficial effects such as, without limitation, an increase in total biomass accumulation, an increase in the percentage of biomass representing carotenoid accumulation, and, in the case of zeaxanthin production, an increase in the hydroxylation of beta-carotene to zeaxanthin. Those of ordinary skill in the art will be able to select without undue experimentation an appropriate rate of increase, an appropriate type of increase (e.g. continuous, discrete steps or a combination of the two), and/or an optimum pH within the selected range to maximize these and/or other beneficial effects.

In some embodiments, the temperature at which inventive cells are cultivated is selected so that production of one or more particular carotenoid compound(s) and/or retinolic compound(s) is adjusted (e.g., so that production of one or more particular compound(s) is increased and/or production of one or more other compound(s) is decreased). In some embodiments, the temperature at which inventive cells are cultivated is selected so that the ratio of one carotenoid compound and/or retinolic compound to another, is adjusted. To give but one example, in some embodiments, a temperature is selected to be sufficiently low that β-carotene levels are reduced and the level of at least one other carotenoid compound(s) (e.g., zeaxanthin) is increased.

In some embodiments, cultures are grown at about pH 5.5, at about pH 7.0, and or at a temperature between about 28-30° C. In some embodiments, it may be desirable to grow inventive modified cells under low pH conditions, in order to minimize growth of other cells. In some embodiments, it will be desirable to grow inventive modified cells under relatively higher temperature conditions in order to slow growth rate and/or increase the ultimate dry cell weight output of carotenoids and/or retinolic compounds. In some embodiments, it will be desirable to grow inventive modified cells under conditions in which the pH in increased (e.g. continuously, in discrete steps, or both) during the course of fermentation (e.g. increased from pH 7.0 to pH 8.0). In some embodiments, it will be desirable to grow inventive modified cells under two or more of these conditions. For example, inventive modified cells can be grown under relatively higher temperature conditions while simultaneously increasing the pH over the course of the fermentation. Those of ordinary skill in the art will be able to select appropriate growth conditions to achieve their experimental, production and/or other cell culture goals.

One advantage provided by the present invention is that, in addition to allowing the production of high levels of carotenoids and/or retinolic compounds, certain embodiments of the present invention allow produced compounds to be readily isolated because they accumulate in the lipid bodies within oleaginous organisms. Methods and systems for isolating lipid bodies have been established for a wide variety of oleaginous organisms (see, for example, U.S. Pat. Nos. 5,164,308; 5,374,657; 5,422,247; 5,550,156; 5,583,019; 6,166,231; 6,541,049; 6,727,373; 6,750,048; and 6,812,001, each of which is incorporated herein by reference in its entirety). In brief, cells are typically recovered from culture, often by spray drying, filtering or centrifugation.

Of course, it is not essential that lipid bodies be specifically isolated in order to collect carotenoid compounds and/or retinolic compounds produced according to the present invention. Any of a variety of approaches can be utilized to isolate and/or purify carotenoids and/or retinolic compounds. Many useful extraction and/or purification procedures for particular carotenoid compounds, and/or for carotenoids generally, are known in the art (see, for example, EP670306, EP719866, U.S. Pat. No. 4,439,629, U.S. Pat. No. 4,680,314, U.S. Pat. No. 5,310,554, U.S. Pat. No. 5,328,845, U.S. Pat. No. 5,356,810, U.S. Pat. No. 5,422,247, U.S. Pat. No. 5,591,343, U.S. Pat. No. 6,166,231, U.S. Pat. No. 6,750,048, U.S. Pat. No. 6,812,001, U.S. Pat. No. 6,818,239, U.S. Pat. No. 7,015,014, US2003/0054070, US2005/0266132, each of which is incorporated herein by reference).

In many typical isolation procedures, cells are disrupted (e.g., mechanically (for example using a bead mill, mashing), enzymatically (e.g. using zymolyase or a β-1,3 glucanase such as Glucanex 200G (Novozyme), chemically (e.g., by exposure to a mild caustic agent such as a detergent or 0.1 N NaOH, for example at room temperature or at elevated temperature), using a reducing agent (e.g. dithiothreitol, β-mercaptoethanol), using high pressure homogenization/shearing, by changing pH, etc. and combinations thereof) to allow access of intracellular carotenoid and/or retinolic compound(s) to an extraction solvent, and are then extracted one or more times. In certain embodiments, cells are disrupted mechanically using a bead mill/mashing at high pressure (e.g. at 25K, 10K-30K, 15K-25K, or 20-25K, pound-force per square inch (psi)). Cells may optionally be concentrated (e.g., to at least about 100 g/L or more, including to at least about 120 g/l, 150 g/l, 175 g/L, 200 g/L or more) and/or dried (e.g., with a spray dryer, double drum dryer (e.g. Blaw Knox double drum dryer), single drum vacuum dryer, etc.), prior to exposure to extraction solvent (and/or prior to disruption or homogenization). Disruption can, of course, be performed prior to and/or during exposure to extraction solvent. After extraction, solvent is typically removed (e.g., by evaporation, for example by application of vacuum, change of temperature, etc.).

In some instances, cells are disrupted and then subjected to supercritical liquid extraction or solvent extraction. Typical liquids or solvents utilized in such extractions include, for example, organic or non-organic liquids or solvents. To give but a few specific examples, such liquids or solvents may include acetone, supercritical fluids (e.g. carbon dioxide, propane, xenon, ethane, propylene, methane, ethylene, ethanol), carbon dioxide, chloroform, ethanol, ethyl acetate, heptane, hexane, isopropanol, methanol, methylene chloride, octane, tetrahydrofuran (THF), cyclohexane, isobutyl acetate, methyl ketone, ethyl ketone, toluene, cyclohexanone, benzene, propylene glycol, vegetable oils (e.g. soybeen soybean oil, rapeseed oil, corn oil, cottonseed oil, canola oil, etc.) and combinations thereof (e.g. hexane:ethyl acetate, combination of a polar and non-polar solvent, combination of an alcohol with either hexane or ethyl acetate). Particular solvents may be selected, for example, based on their ability to solubilize particular carotenoid compounds and/or retinolic compounds, or sets of carotenoid compounds (e.g., all carotenoids) and/or retinolic compounds (e.g., all retinolic compounds), and/or based on regulatory or other considerations (e.g., toxicity, cost, ease of handling, ease of removal, ease of disposal, etc.). For example, more polar carotenoids (e.g., xanthophylls) are known to be extracted more efficiently into extraction solvents with increased polarity. Craft (1992) J. Agric. Food Chem 40, 431-434 which is herein incorporated by reference discusses the relative solubility of two carotenoids, lutein and β-carotene in different solvents.

In some embodiments, combinations of solvents may be utilized. In some embodiments, combinations of a relatively polar solvent (e.g., alcohols, acetone, chloroform, methylene chloride, ethyl acetate, etc.) and a relatively non-polar solvent (e.g., hexane, cyclohexane, oils, etc.) are utilized for extraction. Those of ordinary skill in the art will readily appreciate that different ratios of polar to non-polar solvent may be employed as appropriate in a particular situation. Just to give a few examples, common ratios include 1:1, 2:1, 3:1, 3:2, 95:5, 90:10, 85:15, 80:20, 75:25, 70:30, 65:45, 60:40, 55:45, and 50:50. It will be appreciated that solvents or solvent mixtures of different polarities may be more effective at extracting particular carotenoids (e.g., based on their polarities and/or as a function of other attributes of the host cell material from which they are being extracted). Those of ordinary skill in the art are well able to adjust the overall polarity of the extracting solvent, for instance by adjusting the relative amounts of polar and non-polar solvents in a solvent blend, in order to achieve more efficient extraction.

Extraction may be performed under any of a variety of environmental conditions, including any of a variety of temperatures. For example, extraction may be performed on ice (for example at 4° C., 0° C., less than 0° C.), at room temperature, or at any of a variety of other temperatures. For example, a solvent may be maintained at a selected temperature (e.g., about less than 0, 0, 4, 25, 28, 30, 37, 68, 70, 75, 80, 85, 90, 95, or 100° C.) in order to improve or adjust extraction of a particular desired carotenoid.

Extraction typically yields a crude oil suspension. In some embodiments, the crude oil suspension contains some intact host cells but is at least about 95% free of intact host cells. In some embodiments, the crude oil suspension is at least about 96%, 97%, 98%, or 99% or more free of intact host cells. In some embodiments, the suspension is substantially free of water-soluble cell components (e.g., nucleic acids, cell wall or storage carbohydrates, etc.). In some embodiments, the suspension contains less than about 5%, 4%, 3%, 2%, or 1% or less water-soluble cell components.

Extraction conditions that yield a crude oil suspension will enrich for lipophilic components that accumulate in the lipid bodies within oleaginous organisms. In general, the major components of the lipid bodies consist of triacylglycerols, ergosteryl esters, other steryl esters, free ergosterol, phospholipids, and some proteins, which often function in the synthesis or regulation of the levels of other lipid body components. C16 and C18 (e.g. C16:0, C16:1, C18:0, C18:1, and C18:2) are generally the major fatty acids present in lipid bodies, mainly as components of triacylglycerol and steryl esters.

In some embodiments of the invention, the crude oil suspension contains at least about 2.5% by weight carotenoid compound(s) and/or retinolic compound(s); in some embodiments, the crude oil suspension contains at least about 5% by weight carotenoid compound(s) and/or retinolic compound(s), at least about 10% by weight carotenoid compound(s) and/or retinolic compound(s), at least about 20% by weight carotenoid compound(s) and/or retinolic compound(s), at least about 30% by weight carotenoid compound(s) and/or retinolic compound(s), at least about 40% by weight carotenoid compound(s) and/or retinolic compound(s), or at least about 50% by weight carotenoid compound(s) and/or retinolic compound(s).

The crude oil suspension may optionally be refined as known in the art. Refined oils may be used directly as feed or food additives. Alternatively or additionally, carotenoids and/or retinolic compound can be isolated from the oil using conventional techniques.

Given the sensitivity of carotenoids and retinolic compounds generally to oxidation, many embodiments of the invention employ oxidative stabilizers (e.g., ascorbyl palmitate, tocopherols, vitamin C (e.g. sodium ascorbate), ethoxyquin, vitamin E, BHT, BHA, TBHQ, etc., or combinations thereof) during and/or after carotenoid isolation. Alternatively or additionally, nitrogen or an inert gas can be utilized to purge oxygen from the process lines of any tanks or equipment. Alternatively or additionally, microencapsulation, (for example with a microencapsulation ingredients such as proteins, carbohydrates (e.g. maltodextrin, gum acacia, xanthan gum, starches/sugars like sucrose), or gelatins, or any other substance which creates a physical barrier to air and/or light) may be employed to add a physical barrier to oxidation and/or to improve handling (see, for example, U.S. Patent Applications 2004/0191365 and 2005/0169999). For example, carotenoids and/or retinolic compounds produced according to the present invention may be microencapsulated after isolation during the formulation of commercial products (e.g. pharmaceuticals, food supplements, electro-optic applications, animal feed additives, cosmetics, etc.) to minimize or eliminate oxidation during production, storage, transport, etc.

Extracted carotenoids and/or retinolic compounds may be further isolated and/or purified, for example, by crystallization, washing, recrystallization, and/or other purification strategies. In some embodiments, carotenoid and/or retinolic compound crystals are collected by filtration and/or centrifugation. Isolated or purified carotenoids and/or retinolic compound may be dried and/or formulated for storage, transport, sale, and/or ultimate use. To give but a few specific examples, carotenoids and/or retinolic compounds may be prepared as a water (e.g. cold water) dispersible powder (e.g. 1%-20% carotenoid: microencapsulation ingredient), as a suspension of crystals in oil (e.g., vegetable oil, e.g., about 1%-30%, 5%-30%, 10%-30% w/w), etc.

Uses

Carotenoids and/or retinolic compounds produced according to the present invention can be utilized in any of a variety of applications, for example exploiting their biological or nutritional properties (e.g., anti-oxidant, anti-proliferative, etc.) and/or their pigment properties. For example, according to the present invention, carotenoids may be used in pharmaceuticals (see, for example, Bertram, Nutr. Rev. 57:182, 1999; Singh et al., Oncology 12:1643, 1998; Rock, Pharmacol. Ther. 75:185, 1997; Edge et al, J. Photochem Photobiol 41:189, 1997; U.S. Patent Application 2004/0116514; U.S. Patent Application 2004/0259959), food supplements (see, for example, Koyama et al, J. Photochem Photobiol 9:265, 1991; Bauernfeind, Carotenoids as colorants and vitamin A precursors, Academic Press, NY, 1981; U.S. Patent Application 2004/0115309; U.S. Patent Application 2004/0234579), electro-optic applications, animal feed additives (see, for example, Krinski, Pure Appl. Chem. 66:1003, 1994; Polazza et al., Meth. Enzymol. 213:403, 1992), cosmetics (as anti-oxidants and/or as cosmetics, including fragrances; see for example U.S. Patent Application 2004/0127554), etc. Carotenoids produced in accordance with the present invention may also be used as intermediates in the production of other compounds (e.g., steroids, etc.).

For example, astaxanthin and/or esters thereof may be useful in a variety of pharmaceutical applications and health foods including treatment of inflammatory diseases, asthma, atopic dermatitis, allergies, multiple myeloma, arteriosclerosis, cardiovascular disease, liver disease, cerebrovascular disease, thrombosis, neoangiogenesis-related diseases, including cancer, rheumatism, diabetic retinopathy; macular degeneration and brain disorder, hyperlipidemia, kidney ischemia, diabetes, hypertension, tumor proliferation and metastasis; and metabolic disorders. Additionally, carotenoids and astaxanthin may be useful in the prevention and treatment of fatigue, for improving kidney function in nephropathy from inflammatory diseases, as well as prevention and treatment of other life habit-related diseases. Still further, astaxanthin has been found to play a role as inhibitors of various biological processes, including interleukin inhibitors, phosphodiesterase inhibitors, phospholipase A2 inhibitors, cyclooxygenase-2 inhibitors, matrix metalloproteinase inhibitors, capillary endothelium cell proliferation inhibitors, lipoxygenase inhibitors. See, e.g., Japanese Publication No. 2006022121, published 20060126 (JP Appl No. 2005-301156 filed 20051017); Japanese Publication No. 2006016408, published 20060119 (JP Appl No. 2005-301155 filed 20051017); Japanese Publication No. 2006016409, published 20060119 (JP Appl No. 2005-301157 filed 20051017); Japanese Publication No. 2006016407, published 20060119 (JP Appl No. 2005-301153 filed 20051017); Japanese Publication No. 2006008717, published 20060112 (JP Appl No. 2005-301151 filed 20051017); Japanese Publication No. 2006008716, published 20060112 (JP Appl No. 2005-301150 filed 20051017); Japanese Publication No. 2006008720, published 20060112 (JP Appl No. 2005-301158 filed 20051017); Japanese Publication No. 2006008719, published 20060112 (JP Appl No. 2005-301154 filed 20051017); Japanese Publication No. 2006008718, published 20060112 (JP Appl No. 2005-301152 filed 20051017); Japanese Publication No. 2006008713, published 20060112 (JP Appl No. 2005-301147 filed 20051017); Japanese Publication No. 2006008715, published 20060112 (JP Appl No. 2005-301149 filed 20051017); Japanese Publication No. 2006008714, published 20060112 (JP Appl No. 2005-301148 filed 20051017); and Japanese Publication No. 2006008712, published 20060112 (JP Appl No. 2005-301146 filed 20051017).

As other non-limiting examples, retinolic compounds produced according to the present invention may be used in pharmaceuticals, foodstuff, dietary supplements, electro-optic applications, animal feed additives, cosmetics, etc.

It will be appreciated that, in some embodiments of the invention, carotenoids and/or retinolic compounds produced by manipulated host cells as described herein are incorporated into a final product (e.g., food or feed supplement, pharmaceutical, cosmetic, dye-containing item, etc.) in the context of the host cell. For example, host cells may be lyophilized, freeze dried, frozen or otherwise inactivated, and then whole cells may be incorporated into or used as the final product. The host cell may also be processed prior to incorporation in the product to increase bioavailability (e.g., via lysis). Alternatively or additionally, a final product may incorporate only a portion of the host cell (e.g., fractionated by size, solubility), separated from the whole. For example, in some embodiments of the invention, lipid droplets are isolated from the host cells and are incorporated into or used as the final product. For instance, inventive carotenoid-containing and/or retinolic compound-containing lipid bodies (e.g., from engineered cells, and particularly from engineered fungal cells) may be substituted for the plant oil bodies described in U.S. Pat. No. 6,599,513 (the entire contents of which are hereby incorporated by reference) and incorporated into emulsion or emulsion formulations, as described therein. In other embodiments, the carotenoids and/or retinolic compounds themselves, or individual carotenoid and/or retinolic compounds are isolated and reformulated into a final product.

As stated above, fatty acid and glucoside esters are the predominant carotenoid esters found in nature, whereas additional esters (e.g. with organic acids or inorganic phosphate) can be synthesized to generate useful product forms. For delivery, carotenoid esters can also be formulated as salts of the ester form. See, e.g., US Publication No. 20050096477.

The amount of carotenoid and/or retinolic compound incorporated into a given product may vary dramatically depending on the product, and the particular carotenoid(s) and/or retinolic compound(s) involved. Amounts may range, for example, from less than 0.01% by weight of the product, to more than 1%, 10%, 20%, 30% or more; in some cases the carotenoid and/or retinolic compound may comprise 100% of the product. Thus, amount of carotenoid and/or retinolic compound incorporated into a given product may be, for example, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

In some embodiments of the invention, one or more produced carotenoids and/or retinolic compounds is incorporated into a component of food or feed (e.g., a food supplement). Types of food products into which carotenoids and/or retinolic compounds can be incorporated according to the present invention are not particularly limited, and include beverages such as milk, water, sports drinks, energy drinks, teas, juices, and liquors; confections such as jellies and biscuits; fat-containing foods and beverages such as dairy products; processed food products such as rice and soft rice (or porridge); infant formulas; breakfast cereals; or the like. In some embodiments, one or more produced carotenoids and/or retinolic compounds is incorporated into a dietary supplements, such as for example a multivitamin. In certain embodiments, beta-carotene produced according to the present invention is included in a dietary supplement. In certain embodiments, lutein produced according to the present invention is included in a dietary supplement. In certain embodiments, retinol, retinal, retinyl palmitate, retinyl acetate, and/or retinoic acid produced according to the present invention is included in a dietary supplement. In some embodiments of this aspect of the invention, it may be useful to incorporate the carotenoids and/or retinolic compounds within bodies of edible lipids as it may facilitate incorporation into certain fat-containing food products. Thus, for example, when the edible fungus, Candida utilis is used as a host, its' carotenoid and/or retinolic compound containing lipids may be directly incorporated into a component of food or feed (e.g., a food supplement).

Examples of feedstuffs into which carotenoids and/or retinolic compounds produced in accordance with the present invention may be incorporated include, for instance, pet foods such as cat foods, dog foods and the like, feeds for aquarium fish, cultured fish or crustaceans, etc., feed for farm-raised animals (including livestock and further including fish or crustaceans raised in aquaculture). Food or feed material into which the carotenoid(s) and/or retinolic compound(s) produced in accordance with the present invention is incorporated is preferably palatable to the organism which is the intended recipient. This food or feed material may have any physical properties currently known for a food material (e.g., solid, liquid, soft).

In some embodiments, feedstuffs containing carotenoids and/or retinolic compounds produced in accordance with the present invention are substantially free of intact host cells. For example, feedstuffs of the present invention may be at least about 95% free of intact host cells. In some embodiments, feedstuffs of the present invention are at least about 96%, 97%, 98%, or 99% or more free of intact host cells. Such embodiments are typical when the carotenoids and/or retinolic compounds are highly purified away from the host cell in which they were produced (see section entitled “Production and Isolation of Carotenoids and/or Retinolic Compounds”).

In some embodiments, feedstuffs containing carotenoids and/or retinolic compounds produced in accordance with the present invention are not substantially free of intact host cells. For example, feedstuffs of the present invention may comprise greater than about 95% intact host cells. In certain embodiments, feedstuffs of the present invention comprise greater than about 70%, 75%, 85%, or 90% intact host cells. In certain embodiments, feedstuffs of the present invention comprise nearly intact host cells. For example, feedstuffs of the present invention may comprise greater than about 70%, 75%, 85%, 90%, or 95% nearly intact host cells. As will be appreciated by those of ordinary skill in the art, carotenoid and/or retinolic compound-containing feedstuffs of the present invention that contain intact cells and/or nearly intact cells will have great utility in providing the carotenoids and/or retinolic compounds of interest present in such host cells to an animal. Such embodiments are advantageous when host cells that produce the carotenoids and/or retinolic compounds of interest contain additional vitamins, nutrients, etc. that benefit the animal.

In some embodiments of the invention, one or more produced carotenoids and/or retinolic compounds is incorporated into a cosmetic product. Examples of such cosmetics include, for instance, skin cosmetics (e.g., lotions, emulsions, creams and the like), lipsticks, anti-sunburn cosmetics, makeup cosmetics, fragrances, products for daily use (e.g., toothpastes, mouthwashes, bad breath preventive agents, solid soaps, liquid soaps, shampoos, conditioners), etc.

In some embodiments, one or more produced carotenoids and/or retinolic compounds is incorporated into a pharmaceutical. Examples of such pharmaceuticals include, for instance, various types of tablets, capsules, drinkable agents, troches, gargles, etc. In some embodiments, the pharmaceutical is suitable for topical application. Dosage forms are not particularly limited, and include capsules, oils, granula, granula subtilae, pulveres, tabellae, pilulae, trochisci, or the like. Oils and oil-filled capsules may provide additional advantages both because of their lack of ingredient decomposition during manufacturing, and because inventive carotenoid-containing and/or retinolic compound-containing lipid droplets may be readily incorporated into oil-based formulations.

Pharmaceuticals according to the present invention may be prepared according to techniques established in the art including, for example, the common procedure as described in the United States Pharmacopoeia, for example.

Carotenoids and/or retinolic compounds produced according to the present invention may be incorporated into any pigment-containing product including, for example, fabric, paint, etc. They may also be incorporated into a product which is an environmental indicator, or an instrument such as a biosensor for use as a detection agent.

Carotenoids and/or retinolic compounds produced according to the present invention (whether isolated or in the context of lipid droplets or of cells, e.g., fungal cells) may be incorporated into products as described herein by combinations with any of a variety of agents. For instance, such carotenoids and/or retinolic compounds may be combined with one or more binders or fillers. In some embodiments, inventive products will include one or more chelating agents, pigments, salts, surfactants, moisturizers, viscosity modifiers, thickeners, emollients, fragrances, preservatives, etc., and combinations thereof.

Useful surfactants include, for example, anionic surfactants such as branched and unbranched alkyl and acyl hydrocarbon compounds, sodium dodecyl sulfate (SDS); sodium lauryl sulfate (SLS); sodium lauryl ether sulfate (SLES); sarconisate; fatty alcohol sulfates, including sodium, potassium, ammonium or triethanolamine salts of C₁₀ to C₁₈ saturated or unsaturated forms thereof; ethoxylated fatty alcohol sulfates, including alkyl ether sulfates; alkyl glyceryl ether sulfonate, alpha sulpho fatty acids and esters; fatty acid esters of isethionic acid, including Igepon A; acyl (fatty) N-methyltaurides, including Igepon T; dialkylsulfo succinate esters, including C₈, C₁₀ and C₁₂ forms thereof; Miranot BT also referred to as lauroamphocarboxyglycinate and sodium tridecath sulfate; N-acylated amino acids, such as sodium N-lauroyl sarconisate or gluconate; sodium coconut monoglyceride sulfonate; and fatty acid soaps, including sodium, potassium, DEA or TEA soaps.

Among the cationic surfactants that are useful are monoalkyl trimethyl quartenary salts; dialkyl dimethyl quartenary salts; ethoxylated or propoxylated alkyl quaternary ammonium salts, also referred to in the art as ethoquats and propoquats; cetyl benzylmethylalkyl ammonium chloride; quaternized imidazolines, which are generally prepared by reacting a fat or fatty acid with diethylenetriamine followed by quaternization, and non-fat derived cationic polymers such as the cellulosic polymer, Polymer JR (Union Carbide).

Further useful cationic surfactants include lauryl trimethyl ammonium chloride; cetyl pyridinium chloride; and alkyltrimethylammonium bromide. Cationic surfactants are particularly useful in the formulation of hair care products, such as shampoos, rinses and conditioners.

Useful nonionic surfactants include polyethoxylated compounds and polypropoxylated products. Examples of ethoxylated and propoxylated non-ionic surfactants include ethoxylated anhydrohexitol fatty esters, for example Tween 20; mono- and diethanolamides; Steareth-20, also known as Volpo20; polyethylene glycol fatty esters (PEGs), such as PEG-8-stearate, PEG-8 distearate; block co-polymers, which are essentially combinations of hydrophylic polyethoxy chains and lipophilic polypropoxy chains and generically known as Poloaxamers.

Still other useful non-ionic surfactants include fatty esters of polyglycols or polyhydric alcohols, such as mono and diglyceride esters; mono- and di-ethylene glycol esters; diethylene glycol esters; sorbitol esters also referred to as Spans; sucrose esters; glucose esters; sorbitan monooleate, also referred to as Span80; glyceryl monostearate; and sorbitan monolaurate, Span20 or Arlacel 20.

Yet other useful nonionic surfactants include polyethylene oxide condensates of alkyl phenols and polyhydroxy fatty acid amide surfactants which may be prepared as for example disclosed in U.S. Pat. No. 2,965,576.

Examples of amphoteric surfactants which can be used in accordance with the present invention include betaines, which can be prepared by reacting an alkyldimethyl tertiary amine, for example lauryl dimethylamine with chloroacetic acid. Betaines and betaine derivatives include higher alkyl betaine derivatives including coco dimethyl carboxymethyl betaine; sulfopropyl betaine; alkyl amido betaines; and cocoamido propyl betaine. Sulfosultaines which may be used include for example, cocoamidopropyl hydroxy sultaine. Still other amphoteric surfactants include imidazoline derivatives and include the products sold under the trade name “Miranol” described in U.S. Pat. No. 2,528,378 which is incorporated herein by reference in its entirety. Still other amphoterics include phosphates for example, cocamidopropyl PG-dimonium chloride phosphate and alkyldimethyl amine oxides.

Suitable moisturizers include, for example, polyhydroxy alcohols, including butylene glycol, hexylene glycol, propylene glycol, sorbitol and the like; lactic acid and lactate salts, such as sodium or ammonium salts; C₃ and C₆ diols and triols including hexylene glycol, 1,4 dihydroxyhexane, 1,2,6-hexane triol; aloe vera in any of its forms, for example aloe vera gel; sugars and starches; sugar and starch derivatives, for example alkoxylated glucose; hyaluronic acid; lactamide monoethanolamine; acetamide monoethanolamine; glycolic acid; alpha and beta hydroxy acids (e.g. lactic, glycolic salicylic acid); glycerine; pantheol; urea; vaseline; natural oils; oils and waxes (see: the emollients section herein) and mixtures thereof.)

Viscosity modifiers that may be used in accordance with the present invention include, for example, cetyl alcohol; glycerol, polyethylene glycol (PEG); PEG-stearate; and/or Keltrol.

Appropriate thickeners for use in inventive products include, for example, gelling agents such as cellulose and derivatives; Carbopol and derivatives; carob; carregeenans and derivatives; xanthane gum; sclerane gum; long chain alkanolamides; bentone and derivatives; Kaolin USP; Veegum Ultra; Green Clay; Bentonite NFBC; etc.

Suitable emollients include, for example, natural oils, esters, silicone oils, polyunsaturated fatty acids (PUFAs), lanoline and its derivatives and petrochemicals.

Natural oils which may be used in accordance with the present invention may be obtained from sesame; soybean; apricot kernel; palm; peanut; safflower; coconut; olive; cocoa butter; palm kernel; shea butter; sunflower; almond; avocado; borage; carnauba; hazel nut; castor; cotton seed; evening primrose; orange roughy; rapeseed; rice bran; walnut; wheat germ; peach kernel; babassu; mango seed; black current seed; jojoba; macademia nut; sea buckthorn; sasquana; tsubaki; mallow; meadowfoam seed; coffee; emu; mink; grape seed; thistle; tea tree; pumpkin seed; kukui nut; and mixtures thereof.

Esters which may be used include, for example, C₈-C₃₀ alkyl esters of C₈-C₃₀ carboxylic acids; C₁-C₆ diol monoesters and diesters of C₈-C₃₀ carboxylic acids; C₁₀-C₂₀ alcohol monosorbitan esters, C₁₀-C₂₀ alcohol sorbitan di- and tri-esters; C₁₀-C₂₀ alcohol sucrose mono-, di-, and tri-esters and C₁₀-C₂₀ fatty alcohol esters of C₂-C₆ 2-hydroxy acids and mixtures thereof. Examples of these materials include isopropyl palmitate; isopropyl myristate; isopropyl isononate; C₁₂/C14 benzoate ester (also known as Finesolve); sorbitan palmitate, sorbitan oleate; sucrose palmitate; sucrose oleate; isostearyl lactate; sorbitan laurate; lauryl pyrrolidone carboxylic acid; panthenyl triacetate; and mixtures thereof.

Further useful emollients include silicone oils, including non-volatile and volatile silicones. Examples of silicone oils that may be used in the compositions of the present invention are dimethicone; cyclomethycone; dimethycone-copolyol; aminofunctional silicones; phenyl modified silicones; alkyl modified silicones; dimethyl and diethyl polysiloxane; mixed C₁-C₃₀ alkyl polysiloxane; and mixtures thereof. Additionally useful silicones are described in U.S. Pat. No. 5,011,681 to Ciotti et al., incorporated by reference herein.

A yet further useful group of emollients includes lanoline and lanoline derivatives, for example lanoline esters.

Petrochemicals which may be used as emollients in the compositions of the present invention include mineral oil; petrolatum; isohexdecane; permethyl 101; isododecanol; C₁₁-C₁₂ Isoparrafin, also known as Isopar H.

Among the waxes which may be included in inventive products are animal waxes such as beeswax; plant waxes such as carnauba wax, candelilla wax, ouricurry wax, Japan wax or waxes from cork fibres or sugar cane. Mineral waxes, for example paraffin wax, lignite wax, microcrystalline waxes or ozokerites and synthetic waxes may also be included.

Exemplary fragrances for use in inventive products include, for instance, linear and cyclic alkenes (i.e. terpenes); primary, secondary and tertiary alcohols; ethers; esters; ketones; nitrites; and saturated and unsaturated aldehydes; etc.

Examples of synthetic fragrances that may be used in accordance with the present invention include without limitation acetanisole; acetophenone; acetyl cedrene; methyl nonyl acetaldehyde; musk anbrette; heliotropin; citronellol; sandella; methoxycitranellal; hydroxycitranellal; phenyl ethyl acetate; phenylethylisobutarate; gamma methyl ionone; geraniol; anethole; benzaldehyde; benzyl acetate; benzyl salicate; linalool; cinnamic alcohol; phenyl acetaldehyde; amyl cinnamic aldehyde; caphore; p-tertiary butyl cyclohexyl acetate; citral; cinnamyl acetate; citral diethyl acetal; coumarin; ethylene brasslate; eugenol; l-menthol; vanillin; etc.

Examples of natural fragrances of use herein include without limitation lavandin; heliotropin; sandlewood oil; oak moss; pathouly; ambergris tincture; ambrette seed absolute; angelic root oil; bergamont oil; benzoin Siam resin; buchu leaf oil; cassia oil; cedarwood oil; cassia oil; castoreum; civet absolute; chamomile oil; geranium oil; lemon oil; lavender oil; Ylang Ylang oil; etc.

A list of generally used fragrance materials can be found in various reference sources, for example, “Perfume and Flavor Chemicals”, Vols. I and II; Steffen Arctander Allured Pub. Co. (1994) and “Perfumes: Art, Science and Technology”; Muller, P. M. and Lamparsky, D., Blackie Academic and Professional (1994) both incorporated herein by reference.

Suitable preservatives include, among others, (e.g., sodium metabisulfite; Glydant Plus; Phenonip; methylparaben; Germall 115; Germaben II; phytic acid; sodium lauryl sulfate (SLS); sodium lauryl ether sulfate (SLES); Neolone; Kathon; Euxyl and combinations thereof), anti-oxidants (e.g., butylated hydroxytoluened (BHT); butylated hydroxyanisol (BHA); ascorbic acid (vitamin C); tocopherol; tocopherol acetate; phytic acid; citric acid; pro-vitamin A.

In some embodiments, inventive products will comprise an emulsion (e.g., containing inventive lipid bodies), and may include one or more emulsifying agents (e.g., Arlacel, such as Alacel 165; Glucamate; and combinations thereof) and/or emulsion stabilizing agents.

In some embodiments, inventive products will include one or more biologically active agents other than the carotenoid(s). To give but a few examples, inventive cosmetic or pharmaceutical products may include one or more biologically active agents such as, for example, sunscreen actives, anti-wrinkle actives, anti-aging actives, whitening actives, bleaching actives, sunless tanning actives, anti-microbial actives, anti-acne actives, anti-psoriasis actices, anti-eczema actives, antioxidants, anesthetics, vitamins, protein actives, etc.

EXEMPLIFICATION

Table 26 below describes certain Yarrowia lipolytica strains used in the following exemplification:

TABLE 26 Yarrowia lipolytica strains. NRRL Y-1095 Wild type diploid ATCC76861 MATB ura2-21 lyc1-5 LYS1-5B ATCC76982 MATB ade1 leu2-35 lyc1-5 xpr2 ATCC201249 MATA ura3-302 leu2-270 lys8- 11 PEX17-HA MF346 MATA ura2-21 ATCC76861 × ATCC201249 MF350 MATB ura2-21 leu2-35 ade1 ATCC76982 × MF346

(The genotypes at LYC1, LYS1, XPR2, and PEX17 were not determined in crosses nor verified for ATCC strains.)

All basic molecular biology and DNA manipulation procedures described herein are generally performed according to Sambrook et al. or Ausubel et al. (Sambrook J, Fritsch E F, Maniatis T (eds). 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press: New York; Ausubel F M, Brent R, Kingston R E, Moore D D, Seidman J G, Smith J A, Struhl K (eds). 1998. Current Protocols in Molecular Biology. Wiley: New York).

Example 1 Production of Plasmids for Carotenoid Strain Construction

Plasmids were generated for construction of carotenoid producing strains. The following subparts describe production of plasmids encoding carotenogenic polypeptides. Plasmids used in these studies and details of their construction are described in Table 27. Additional plasmid construction details and descriptions of their use are found in the text of the relevant subsection. All PCR amplifications used NRRL Y-1095 genomic DNA as template unless otherwise specified. The URA5 gene described below is allelic with the ura2-21 auxotrophy above. The GPD1 and TEF1 promoters are from Y. lipolytica as is the XPR2 terminator.

GGS1 is the gene encoding the Y. lipolytica gene encoding geranylgeranylpyrophosphate synthase. The nucleic acid coding sequence, and encoded Ggs1 protein of pMB4591 and pMB4683 are as follows:

(SED ID NO: 1) atggattataacagcgcggatttcaaggagatatggggcaaggccgccga caccgcgctgctgggaccgtacaactacctcgccaacaaccggggccaca acatcagagaacacttgatcgcagcgttcggagcggttatcaaggtggac aagagcgatctcgagaccatttcgcacatcaccaagattttgcataactc gtcgctgcttgttgatgacgtggaagacaactcgatgctccgacgaggcc tgccggcagcccattgtctgtttggagtcccccaaaccatcaactccgcc aactacatgtactttgtggctctgcaggaggtgctcaagctcaagtctta tgatgccgtctccattttcaccgaggaaatgatcaacttgcatagaggtc agggtatggatctctactggagagaaacactcacttgcccctcggaagac gagtatctggagatggtggtgcacaagaccggtggactgtttcggctggc tctgagacttatgctgtcggtggcatcgaaacaggaggaccatgaaaaga tcaactttgatctcacacaccttaccgacacactgggagtcatttaccag attctggatgattacctcaacctgcagtccacggaattgaccgagaacaa gggattctgcgaagatatcagcgaaggaaagttttcgtttccgctgattc acagcatacgcaccaacccggataaccacgagattctcaacattctcaaa cagcgaacaagcgacgcttcactcaaaaagtacgccgtggactacatgag aacagaaaccaagagtttcgactactgcctcaagaggatacaggccatgt cactcaaggcaagttcgtacattgatgatctagcagcagctggccacgat gtctccaagctacgagccattttgcattattttgtgtccacctctgactg tgaggagagaaagtactttgaggatgcgcagtga (SEQ ID NO: 2) mdynsadfkeiwgkaadtallgpynylannrghnirehliaafgavikvd ksdletishitkilhnssllvddvednsmlrrglpaahclfgvpqtinsa nymyfvalqevlklksydavsifteeminlhrgqgmdlywretltcpsed eylemvvhktgglfrlalrlmlsvaskqedhekinfdlthltdtlgviyq ilddylnlqsteltenkgfcedisegkfsfplihsirtnpdnheilnilk qrtsdaslkkyavdymrtetksfdyclkriqamslkassyiddlaaaghd vsklrailhyfvstsdceerkyfedaq

TABLE 27 Plasmids Oligos Plasmid Backbone Insert or source pMB4529 PCR2.1 3.4 kb ADE1 PCR MO4475 & product MO4476 pMB4534 PCR2.1 2.1 kb LEU2 PCR MO4477 & product MO4478 pMB4535 PCR2.1 1.2 kb URA5 PCR MO4471 & product MO4472 pMB4589 pMB4535 1.2 kb GPD1 promoter MO4568 & (KpnI + SpeI) (KpnI + NotI); 0.14 kb MO4591; XPR2 terminator (NotI + MO4566 & SpeI) MO4593 pMB4590 pMB4535 0.4 kb TEF1 promoter MO4571 & (KpnI + SpeI) (KpnI + NotI); 0.14 kb MO4592; XPR2 terminator (NotI + MO4566 & SpeI) MO4593 pMB4591 pMB4590 1.0 kb GGS1 ORF (XbaI MO4534 & (NheI + MluI) + MluI) MO4544 pMB4597 pMB4534 GPD1 promoter & XPR2 From (Acc65I +SpeI) terminator (Acc65I + pMB4589 SpeI) pMB4603 pMB4597 Residual backbone From (RsrII + MluI) & TEF1 promoter (RsrII + pMB4590 MluI) pMB4616 pMB4529 Residual backbone From (RsrII + SpeI) & GPD1 promoter & pMB4589 XPR2 terminator (RsrII + SpeI) pMB4629 pMB4616 Residual backbone From (RsrII + MluI) & TEF1 promoter (RsrII + pMB4590 MluI) pMB4631 pMB4603 1.2 kb GPD1 promoter MO4568 & (KpnI + NheI) (KpnI + NheI); MO4659 pMB4628 pMB4603 carRP See 1A pMB4637 pMB4629 1.5 kb hmg1^(trunc) ORF See 1D (NheI + MluI) (XbaI + MluI) pMB4714 pMB4691 1.5 kb hmg1^(trunc) ORF See 1D (NheI + MluI) (XbaI + MluI) pMB4638 pMB4629 carB(i⁻) See 1B pMB4660 pMB4638 (+URA 3) carB(i⁻) See 1C pMB4662 pMB4631 1.8 kb URA3 fragment See 1C (SpeI + XhoI) (SpeI + BsaI pMB4683 pMB4662 (Acc65I + 1.4 kb tef1p-GGS1 From MluI) fragment (Acc65I + pMB4591 MluI) pMB4692 pMB4662 (Acc65I + 0.4 kb TEF1 promoter See 1E MluI) (Acc65I + NheI); 0.55 kb crtZ ORF (XbaI + MluI) pMB4698 pMB4629 0.9 kb crtW ORF (XbaI + See 1F (NheI + MluI) MluI) pMB4599 pBluescriptSKII- 1.9 kb carRP gene See 1A (EcoRV) pMB4606 pBluescriptSKII- 1.9 kb carB gene See 1A (EcoRV) pMB4613 pMB4599 (Acc65I + carRP(i⁻) See 1A PpuMI) pMB4619 pBluescriptSKII- carB(i⁻) See 1A (BamHI + Acc65I)) pMB4705 pMB4603 carRP(i⁻) See 1A (NheI + MluI) pMB4691 pMB4662 0.4 kb TEF1 promoter From (Acc65I + MluI) (Acc65I + MluI) pMB4629 pMB4751 pMB4691 0.75 kb YALI0D12903g Inserted promoter + 0.45 Y. between lipoltyica HIS3 TEF1p/ terminator XPRt and URA3 pMB4719 pMB4691 crtZ (E. litoralis) See 1J (NheI + MluI) pMB4778 pMB4751 crtZ (P. bermudensis) See 1I (NheI + MluI) pMB4741 pMB4629 crtW (Aurantimonas) See 1G (NheI + MluI) pMB4735 pMB4629 crtW (P. bermudensis) See 1H (NheI + MluI) pMB4812 pMB4603 al-2 (N. crassa) See 1K (NheI + MluI) pMB4846 pMB4691 crtZ (Erythrobacter sp. See 1L (NheI + MluI) NAP1) pMB4835 pMB4691 S. alaskensis) See 1M (NheI + MluI) pMB4845 pMB4691 crtZ (R. biformata) See 1N (NheI + MluI) pMB4837 pMB4691 crtZ (X. autrophicus) See 1O (NheI + MluI) pMB4850 pMB4691 crtZ (P. putida) See 1P (NheI + MluI)

Certain oligonucleotides referred to in Table 27 above are as follows:

MO4471 (SEQ ID NO: 3) 5′-CTGGGTGACCTGGAAGCCTT MO4472 (SEQ ID NO: 4) 5′-AAGATCAATCCGTAGAAGTTCAG MO4475 (SEQ ID NO: 5) 5′-AAGCGATTACAATCTTCCTTTGG MO4476 (SEQ ID NO: 6) 5′-CCAGTCCATCAACTCAGTCTCA MO4477 (SEQ ID NO: 7) 5′-GCATTGCTTATTACGAAGACTAC MO4478 (SEQ ID NO: 8) 5′-CCACTGTCCTCCACTACAAACAC MO4534 (SEQ ID NO: 9) 5′-CACAAACGCGTTCACTGCGCATCCTCAAAGT MO4544 (SEQ ID NO: 10) 5′-CACAATCTAGACACAAATGGATTATAACAGCGCGGAT MO4566 (SEQ ID NO: 11) 5′-CACAAACTAGTTTGCCACCTACAAGCCAGAT MO4568 (SEQ ID NO: 12) 5′-CACAAGGTACCAATGTGAAAGTGCGCGTGAT MO4571 (SEQ ID NO: 13) 5′-CACAAGGTACCAGAGACCGGGTTGGCGG MO4591 (SEQ ID NO: 14) 5′-CACAAGCGGCCGCGCTAGCATGGGGATCGATCTCTTATAT MO4592 (SEQ ID NO: 15) 5′-CACAAGCGGCCGCGCTAGCGAATGATTCTTATACTCAGAAG MO4593 (SEQ ID NO: 16) 5′-CACAAGCGGCCGCACGCGTGCAATTAACAGATAGTTTGCC MO4659 (SEQ ID NO: 17) 5′-CACAAGCTAGCTGGGGATGCGATCTCTTATATC 1A: Production of pMB4628 (tef1p-carRP LEU2) and pMB4705 (tef1p-carRP[i⁻] LEU2) Encoding Phytoene Synthase/Lycopene Cyclase:

Intron-containing carRP was amplified from M. circinelloides (ATCC 90680) genomic DNA using MO4525 and MO4541:

MO4525 (SEQ ID NO: 18) 5′-CACAAACGCGTTTAAATGGTATTTAGATTTCTCATT MO4541 (SEQ ID NO: 19) 5′-CACAATCTAGACACAAATGCTGCTCACCTACATGGA and the resulting 1.9 kb fragment was phosphorylated with T4 polynucleotide kinase. The resulting fragment was blunt-end ligated into pBluescriptSKII-cleaved with EcoRV, yielding pMB4599. The 1.9 kb XbaI-MluI fragment from pMB4599 was inserted into NheI- and MluI-cleaved pMB4603, yielding pMB4628. The intron containing nucleic acid coding sequence, and encoded CarRP protein (assuming correctly predicted splicing) of pMB4628 are as follows:

(SEQ ID NO: 20) atgctgctcacctacatggaagtccacctctactacacgctgcctgtgct gggcgtcctgtcctggctgtcgcggccgtactacacagccaccgatgcgc tcaaattcaaatttctgacactggttgccttcacgaccgcctccgcctgg gacaactacattgtctaccacaaggcgtggtcctactgccccacctgcgt caccgctgtcattggctacgtgcccttggaggagtacatgttcttcatca tcatgactctgttgaccgtggcattcaccaatctggtgatgcgctggcac ctgcacagcttctttatcaggcctgaaacgcccgtcatgcagtccgtcct ggtccgtcttgtccccataacagccttattaatcactgcatacaaggctt gggtaagcaaacaaacaaatgatgtgccgcatcgcattttaatattaacc attgcatacacagcatttggcggtccctggaaagccactgttctacggat catgcattttgtggtacgcctgtccggttttggccttattgtggtttggt gctggcgagtacatgatgcgtcgtccgctggcggtgctcgtctccattgc gctgcccacgctgtttctctgctgggtcgatgtcgtcgctattggcgccg gcacatgggacatttcgctggccacaagcaccggcaagttcgtcgtgccc cacctgcccgtggaggaattcatgttctttgcgctaattaataccgtttt ggtatttggtacgtgtgcgatcgatcgcacgatggcgatcctccacctgt tcaaaaacaagagtccttatcagcgcccataccagcacagcaagtcgttc ctccaccagatcctcgagatgacctgggccttctgtttacccgaccaagt gctgcattcagacacattccacgacctgtccgtcagctgggacatcctgc gcaaggcctccaagtccttttacacggcctctgctgtctttcccggcgac gtgcgccaagagctcggtgtgctatacgccttttgcagagccacggacga tctctgcgacaacgagcaggtccctgtgcagacgcgaaaggagcagctga tactgacacatcagttcgtcagcgatctgtttggccaaaagacaagcgcg ccgactgccattgactgggacttttacaacgaccaactgcctgcctcgtg catctctgccttcaagtcgttcacccgtttgcgccatgtgctggaagctg gagccatcaaggaactgctcgacgggtacaagtgggatttggagcgtcgc tccatcagggatcaggaggatctcagatattactcagcttgtgtcgccag cagtgttggtgaaatgtgcactcgcatcatactggcccacgccgacaagc ccgcctcccgccagcaaacacagtggatcattcagcgtgcgcgtgaaatg ggtctggtactccaatatacaaacattgcaagagacattgtcaccgacag cgaggaactgggcagatgctacctgcctcaggattggcttaccgagaagg aggtggcgctgattcaaggcggccttgcccgagaaattggcgaggagcga ttgctctcactgtcgcatcgcctcatctaccaggcagacgagctcatggt ggttgccaacaagggcatcgacaagctgcccagccattgtcaaggcggcg tgcgtgcggcctgcaacgtctatgcttccattggcaccaagctcaagtct tacaagcaccactatcccagcagagcacatgtcggcaattcgaaacgagt ggaaattgctcttcttagcgtatacaacctttacaccgcgccaattgcga ctagtagtaccacacattgcagacagggaaaaatgagaaatctaaatacc atttaa (SEQ ID NO: 21) mlltymevhlyytlpvlgvlswlsrpyytatdalkfkfltlvafttasaw dnyivyhkawsycptcvtavigyvpleeymffiimtlltvaftnlvmrwh lhsffirpetpvmqsvlvrlvpitallitaykawhlavpgkplfygscil wyacpvlallwfgageymmrrplavlvsialptlflcwvdvvaigagtwd islatstgkfvvphlpveefmffalintvlvfgtcaidrtmailhlfknk spyqrpyqhsksflhqilemtwafclpdqvlhsdtfhdlsvswdilrkas ksfytasavfpgdvrqelgvlyafcratddlcdneqvpvqtrkeqlilth qfvsdlfgqktsaptaidwdfyndqlpascisafksftrlrhvleagaik elldgykwdlerrsirdqedlryysacvassvgemctriilahadkpasr qqtqwiiqraremglvlqytniardivtdseelgrcylpqdwltekeval iqgglareigeerllslshrliyqadelmvvankgidklpshcqggvraa cnvyasigtklksykhhypsrahvgnskrveiallsvynlytapiatsst thcrqgkmrnlnti

Alternatively, pMB4599 was also used as a template for PCR amplification using MO4318, MO4643, MO4644, and MO4639:

MO4318 (SEQ ID NO: 22) 5′-GTAAAACGACGGCCAGT MO4643 (SEQ ID NO: 23) 5′-CACACGGTCTCATGCCAAGCCTTGTATGCAGTGATTAA MO4639 (SEQ ID NO: 24) 5′-CCACTGTGTTTGCTGGCGG MO4644 (SEQ ID NO: 25) 5′-CACACGGTCTCTGGCATTTGGCGGTCCCTGGAAA producing fragments of 0.5 and 0.95 kb, that were subsequently cleaved with Acc65I and BsaI, and BsaI and PpuMI, respectively. These fragments were ligated to pMB4599 that had been digested with Acc65I and PpuMI, yielding pMB4613, harboring intronless carRP. The 1.85 kb XbaI-MluI fragment from pMB4613 was inserted into NheI- and MluI-cleaved pMB4603 to yield pMB4705.

The intronless nucleic acid coding sequence of pMB4705 is as follows, and encodes the same CarRP protein as above:

(SEQ ID NO: 26) atgctgctcacctacatggaagtccacctctactacacgctgcctgtgct gggcgtcctgtcctggctgtcgcggccgtactacacagccaccgatgcgc tcaaattcaaatttctgacactggttgccttcacgaccgcctccgcctgg gacaactacattgtctaccacaaggcgtggtcctactgccccacctgcgt caccgctgtcattggctacgtgcccttggaggagtacatgttcttcatca tcatgactctgttgaccgtggcattcaccaatctggtgatgcgctggcac ctgcacagcttctttatcaggcctgaaacgcccgtcatgcagtccgtcct ggtccgtcttgtccccataacagccttattaatcactgcatacaaggctt ggcatttggcggtccctggaaagccactgttctacggatcatgcattttg tggtacgcctgtccggttttggccttattgtggtttggtgctggcgagta catgatgcgtcgtccgctggcggtgctcgtctccattgcgctgcccacgc tgtttctctgctgggtcgatgtcgtcgctattggcgccggcacatgggac atttcgctggccacaagcaccggcaagttcgtcgtgccccacctgcccgt ggaggaattcatgttctttgcgctaattaataccgttttggtatttggta cgtgtgcgatcgatcgcacgatggcgatcctccacctgttcaaaaacaag agtccttatcagcgcccataccagcacagcaagtcgttcctccaccagat cctcgagatgacctgggccttctgtttacccgaccaagtgctgcattcag acacattccacgacctgtccgtcagctgggacatcctgcgcaaggcctcc aagtccttttacacggcctctgctgtctttcccggcgacgtgcgccaaga gctcggtgtgctatacgccttttgcagagccacggacgatctctgcgaca acgagcaggtccctgtgcagacgcgaaaggagcagctgatactgacacat cagttcgtcagcgatctgtttggccaaaagacaagcgcgccgactgccat tgactgggacttttacaacgaccaactgcctgcctcgtgcatctctgcct tcaagtcgttcacccgtttgcgccatgtgctggaagctggagccatcaag gaactgctcgacgggtacaagtgggatttggagcgtcgctccatcaggga tcaggaggatctcagatattactcagcttgtgtcgccagcagtgttggtg aaatgtgcactcgcatcatactggcccacgccgacaagcccgcctcccgc cagcaaacacagtggatcattcagcgtgcgcgtgaaatgggtctggtact ccaatatacaaacattgcaagagacattgtcaccgacagcgaggaactgg gcagatgctacctgcctcaggattggcttaccgagaaggaggtggcgctg attcaaggcggccttgcccgagaaattggcgaggagcgattgctctcact gtcgcatcgcctcatctaccaggcagacgagctcatggtggttgccaaca agggcatcgacaagctgcccagccattgtcaaggcggcgtgcgtgcggcc tgcaacgtctatgcttccattggcaccaagctcaagtcttacaagcacca ctatcccagcagagcacatgtcggcaattcgaaacgagtggaaattgctc ttcttagcgtatacaacctttacaccgcgccaattgcgactagtagtacc acacattgcagacagggaaaaatgagaaatctaaataccatttaa 1B: Production of pMB4638 (tef1p-carB ADE1), Encoding Phytoene Dehydrogenase:

Intron-containing carB was amplified from M. circinelloides (ATCC 90680) genomic DNA using MO4530 and MO4542:

MO4530 (SEQ ID NO: 27) 5′-CACAAACGCGTTTAAATGACATTAGAGTTATGAAC MO4542 (SEQ ID NO: 28) 5′-CACAATCTAGACACAAATGTCCAAGAAACACATTGTC and the resulting 1.9 kb fragment was phosphorylated with T4 polynucleotide kinase and blunt-end ligated into pBS-SKII-cleaved with EcoRV, yielding pMB4606. pMB4606 was then used as a template for PCR amplification using MO4318 and MO4648, and MO4646 and MO4647, and MO4343 and MO4645:

MO4318 (SEQ ID NO: 29) 5′-GTAAAACGACGGCCAGT MO4648 (SEQ ID NO: 30) 5′-CACAAGGTCTCAAGCACGCATCCCGGAACTG MO4646 (SEQ ID NO: 31) 5′-CACACGGTCTCAGGCATGTCGCCCTACGATGC MO4647 (SEQ ID NO: 32) 5′-CACACGGTCTCATGCTTGCACCCACAAAGAATAGG MO4343 (SEQ ID NO: 33) 5′-CAGGAAACAGCTATGAC MO4645 (SEQ ID NO: 34) 5′-CACACGGTCTCTTGCCCATATACATGGTCTGAAACG producing fragments of 0.4 and 0.85 and 0.7 kb, that were subsequently cleaved with Acc65I and BsaI, and BsaI, and BsaI and BamHI, respectively. These fragments were ligated to pBS-SKII- that had been cut with Acc65I and BamHI, yielding pMB4619, harboring intronless carB. The 1.75 kb XbaI-MluI fragment from pMB4619 was inserted into NheI- and MluI-cleaved pMB4629, yielding pMB4638. The resulting nucleic acid coding sequence and encoded CarB protein of pMB4638 are as follows:

(SEQ ID NO: 35) atgtccaagaaacacattgtcattatcggtgctggcgtgggtggcacggc tacagctgctcgtttggcccgcgaaggcttcaaggtcactgtggtggaga aaaacgactttggtggcggccgctgctccttgatccatcaccagggccat cgctttgatcagggcccgtcgctctacctgatgcccaagtactttgagga cgcctttgccgatctggacgagcgcattcaagaccacctggagctgctgc gatgcgacaacaactacaaggtgcactttgacgacggtgagtcgatccag ctgtcgtctgacttgacacgcatgaaggctgaattggaccgcgtggaggg cccccttggttttggccgattcctggatttcatgaaagagacacacatcc actacgaaagcggcaccctgattgcgctcaagaagaatttcgaatccatc tgggacctgattcgcatcaagtacgctccagagatctttcgcttgcacct gtttggcaagatctacgaccgcgcttccaagtacttcaagaccaagaaga tgcgcatggcattcacgtttcagaccatgtatatgggcatgtcgccctac gatgcgcctgctgtctacagcctgttgcagtacaccgagttcgctgaagg catctggtatccccgtggcggcttcaacatggtggttcagaagctagagg cgattgcaaagcaaaagtacgatgccgagtttatctacaatgcgcctgtt gccaagattaacaccgatgatgccaccaaacaagtgacaggtgtaacctt ggaaaatggccacatcatcgatgccgatgcggttgtgtgtaacgcagatc tggtctatgcttatcacaatctgttgcctccctgccgatggacgcaaaac acactggcttccaagaaattgacgtcttcttccatttccttctactggtc catgtccaccaaggtgcctcaattggacgtgcacaacatctttttggccg aggcttatcaggagagctttgacgaaatcttcaaggactttggcctgcct tctgaagcctccttctacgtcaatgtgccctctcgcatcgatccttctgc tgctcccgacggcaaggactctgtcattgtcttggtgcctattggtcata tgaagagcaagacgggcgatgcttccaccgagaactacccggccatggtg gacaaggcacgcaagatggtgctggctgtgattgagcgtcgtctgggcat gtcgaatttcgccgacttgattgagcatgagcaagtcaatgatcccgctg tatggcagagcaagttcaatctgtggagaggctcaattctgggtttgtct catgatgtgcttcaggtgctgtggttccgtcccagcacaaaggattctac cggtcgttatgataacctattctttgtgggtgcaagcacgcatcccggaa ctggtgttcccattgtccttgcaggaagcaagctcacctctgaccaagtt gtcaagagctttggaaagacgcccaagccaagaaagatcgagatggagaa cacgcaagcacctttggaggagcctgatgctgaatcgacattccctgtgt ggttctggttgcgcgctgccttttgggtcatgtttatgttcttttacttc ttccctcaatccaatggccaaacgcccgcatcttttatcaataatttgtt acctgaagtattccgcgttcataactctaatgtcatttaa (SEQ ID NO: 36) mskkhiviigagvggtataarlaregfkvtvvekndfgggrcslihhqgh rfdqgpslylmpkyfedafadlderiqdhlellrcdnnykvhfddgesiq lssdltrmkaeldrvegplgfgrfldfmkethihyesgtlialkknfesi wdlirikyapeifrlhlfgkiydraskyfktkkmrmaftfqtmymgmspy dapavysllqytefaegiwyprggfnmvvqkleaiakqkydaefiynapv akintddatkqvtgvtlenghiidadavvcnadlvyayhnllppcrwtqn tlaskkltsssisfywsmstkvpqldvhniflaeayqesfdeifkdfglp seasfyvnvpsridpsaapdgkdsvivlvpighmksktgdastenypamv dkarkmvlavierrlgmsnfadlieheqvndpavwqskfnlwrgsilgls hdvlqvlwfrpstkdstgrydnlffvgasthpgtgvpivlagskltsdqv vksfgktpkprkiementqapleepdaestfpvwfwlraafwvmfmffyf fpqsngqtpasfinnllpevfrvhnsnvi 1C. Production of pMB4660 (tef1p-carB URA3) Encoding Phytoene Dehydrogenase:

The 4.3 kb XhoI-NotI fragment and the 1.8 kb NotI-SpeI fragment from pMB4638 were ligated to the 1.9 kb BsaI- and SpeI-cleaved URA3 gene generated by PCR amplification of Y. lipolytica genomic DNA using MO4684 and MO4685 to create pMB4660:

MO4684 (SEQ ID NO: 37) 5′-CATTCACTAGTGGTGTGTTCTGTGGAGCATTC MO4685 (SEQ ID NO: 38) 5′-CACACGGTCTCATCGAGGTGTAGTGGTAGTGCAGTG The resulting nucleic acid coding sequence and encoded CarB(i) protein of pMB4660 are as follows:

(SEQ ID NO: 39) atgtccaagaaacacattgtcattatcggtgctggcgtgggtggcacggc tacagctgctcgtttggcccgcgaaggcttcaaggtcactgtggtggaga aaaacgactttggtggcggccgctgctccttgatccatcaccagggccat cgctttgatcagggcccgtcgctctacctgatgcccaagtactttgagga cgcctttgccgatctggacgagcgcattcaagaccacctggagctgctgc gatgcgacaacaactacaaggtgcactttgacgacggtgagtcgatccag ctgtcgtctgacttgacacgcatgaaggctgaattggaccgcgtggaggg cccccttggttttggccgattcctggatttcatgaaagagacacacatcc actacgaaagcggcaccctgattgcgctcaagaagaatttcgaatccatc tgggacctgattcgcatcaagtacgctccagagatctttcgcttgcacct gtttggcaagatctacgaccgcgcttccaagtacttcaagaccaagaaga tgcgcatggcattcacgtttcagaccatgtatatgggcatgtcgccctac gatgcgcctgctgtctacagcctgttgcagtacaccgagttcgctgaagg catctggtatccccgtggcggcttcaacatggtggttcagaagctagagg cgattgcaaagcaaaagtacgatgccgagtttatctacaatgcgcctgtt gccaagattaacaccgatgatgccaccaaacaagtgacaggtgtaacctt ggaaaatggccacatcatcgatgccgatgcggttgtgtgtaacgcagatc tggtctatgcttatcacaatctgttgcctccctgccgatggacgcaaaac acactggcttccaagaaattgacgtcttcttccatttccttctactggtc catgtccaccaaggtgcctcaattggacgtgcacaacattctttttggcc gaggcttatcaggagagctttgacgaaatcttcaaggactttggcctgcc ttctgaagcctccttctacgtcaatgtgccctctcgcatcgatccttctg ctgctcccgacggcaaggactctgtcattgtcttggtgcctattggtcat atgaagagcaagacgggcgatgcttccaccgagaactacccggccatggt ggacaaggcacgcaagatggtgctggctgtgattgagcgtcgtctgggca tgtcgaatttcgccgacttgattgagcatgagcaagtcaatgatcccgct gtatggcagagcaagttcaatctgtggagaggctcaattctgggtttgtc tcatgatgtgcttcaggtgctgtggttccgtcccagcacaaaggattcta ccggtcgttatgataacctattctttgtgggtgcaagcacgcatcccgga actggtgttcccattgtccttgcaggaagcaagctcacctctgaccaagt tgtcaagagctttggaaagacgcccaagccaagaaagatcgagatggaga acacgcaagcacctttggaggagcctgatgctgaatcgacattccctgtg tggttctggttgcgcgctgccttttgggtcatgtttatgttcttttactt cttccctcaatccaatggccaaacgcccgcatcttttatcaataatttgt tacctgaagtattccgcgttcataactctaatgtcatttaa (SEQ ID NO: 40) mskkhiviigagvggtataarlaregfkvtvvekndfgggrcslihhqgh rfdqgpslylmpkyfedafadlderiqdhlellrcdnnykvhfddgesiq lssdltrmkaeldrvegplgfgrfldfmkethihyesgtlialkknfesi wdlirikyapeifrlhlfgkiydraskyfktkkmrmaftfqtmymgmspy dapavysllqytefaegiwyprggfnmvvqkleaiakqkydaefiynapv akintddatkqvtgvtlenghiidadavvcnadlvyayhnllppcrwtqn tlaskkltsssisfywsmstkvpqldvhniflaeayqesfdeifkdfglp seasfyvnvpsridpsaapdgkdsvivlvpighmksktgdastenypamv dkarkmvlavierrlgmsnfadlieheqvndpavwqskfnlwrgsilgls hdvlqvlwfrpstkdstgrydnlffvgasthpgtgvpivlagskltsdqv vksfgktpkprkiementqapleepdaestfpvwfwlraafwvmfmffyf fpqsngqtpasfinnllpevfrvhnsnvi 1D. Production of pMB4637, pMB4714 and pTef-HMG Encoding a Truncated HMG1.

For production of a truncated variant of the HMG-CoA reductase gene, which also encodes a 77 amino acid leader sequence derived from S. cerevisiae, the following oligonucleotides are synthesized:

PRIMER O (SEQ ID NO: 41) 5′-TTCTAGACACAAAAATGGCTGCAGACCAATTGGTGA PRIMER P (SEQ ID NO: 42) 5′-CATTAATTCTTCTAAAGGACGTATTTTCTTATC PRIMER Q (SEQ ID NO: 43) 5′-GTTCTCTGGACGACCTAGAGG MO4658 (SEQ ID NO: 44) 5′-CACACACGCGTACACCTATGACCGTATGCAAAT

Primers O and P are used to amplify a 0.23 kb fragment encoding Met-Ala followed by residues 530 to 604 of the Hmg1 protein of S. cerevisiae, using genomic DNA as template. Primers Q and MO4658 are used to amplify a 1.4 kb fragment encoding the C-terminal 448 residues of the Hmg1 protein of Y. lipolytica, using genomic DNA as template. These fragments are ligated to the appropriate cloning vector, and the resultant plasmids, designated pOP and pQMO4658, are verified by sequencing. The OP fragment is liberated with XbaI and AseI, and the QMO4658 fragment is liberated with MaeI and MluI. These fragments are then ligated to the ADE1 TEF1p expression vector pMB4629 cut with XbaI and MluI to produce pTefHMG.

Alternatively, the native HMG1 gene from Y. lipolytica was amplified without S. cerevisiae sequences using primers MO4658 (described above) and MO4657 (5′-CACACTCTAGACACAAAAATGACCCAGTCTGTGAAGGTGG (SEQ ID NO:45)). The 1.5 kb product was phosphorylated and ligated to pBluescriptSK⁻ that had been cleaved with EcoRV to create pMB4623. The XbaI-MluI fragment containing hmg1^(trunc) was ligated both to NheI-MluI-cleaved MB4629 and to NheI-MluI-cleaved pMB4691 to create pMB4637 and pMB4714, respectively.

The resulting nucleic acid coding sequence and encoded Hmg1^(trunc) protein of pMB4637 and pMB4714 are as follows:

(SEQ ID NO: 46) atgacccagtctgtgaaggtggttgagaagcacgttcctatcgtcattga gaagcccagcgagaaggaggaggacacctcttctgaagactccattgagc tgactgtcggaaagcagcccaagcccgtgaccgagacccgttctctggac gacctagaggctatcatgaaggcaggtaagaccaagcttctggaggacca cgaggttgtcaagctctctctcgagggcaagcttcctttgtatgctcttg agaagcagcttggtgacaacacccgagctgttggcatccgacgatctatc atctcccagcagtctaataccaagactttagagacctcaaagcttcctta cctgcactacgactacgaccgtgtttttggagcctgttgcgagaacgtta ttggttacatgcctctccccgttggtgttgctggccccatgaacattgat ggcaagaactaccacattcctatggccaccactgagggttgtcttgttgc ctcaaccatgcgaggttgcaaggccatcaacgccggtggcggtgttacca ctgtgcttactcaggacggtatgacacgaggtccttgtgtttccttcccc tctctcaagcgggctggagccgctaagatctggcttgattccgaggaggg tctcaagtccatgcgaaaggccttcaactccacctctcgatttgctcgtc tccagtctcttcactctacccttgctggtaacctgctgtttattcgattc cgaaccaccactggtgatgccatgggcatgaacatgatctccaagggcgt cgaacactctctggccgtcatggtcaaggagtacggcttccctgatatgg acattgtgtctgtctcgggtaactactgcactgacaagaagcccgcagcg atcaactggatcgaaggccgaggcaagagtgttgttgccgaagccaccat ccctgctcacattgtcaagtctgttctcaaaagtgaggttgacgctcttg ttgagctcaacatcagcaagaatctgatcggtagtgccatggctggctct gtgggaggtttcaatgcacacgccgcaaacctggtgaccgccatctacct tgccactggccaggatcctgctcagaatgtcgagtcttccaactgcatca cgctgatgagcaacgtcgacggtaacctgctcatctccgtttccatgcct tctatcgaggtcggtaccattggtggaggtactattttggagccccaggg ggctatgctggagatgcttggcgtgcgaggtcctcacatcgagacccccg gtgccaacgcccaacagcttgctcgcatcattgcttctggagttcttgca gcggagctttcgctgtgttctgctcttgctgccggccatcttgtgcaaag tcatatgacccacaaccggtcccaggctcctactccggccaagcagtctc aggccgatctgcagcgtctacaaaacggttcgaatatttgcatacggtca tag (SEQ ID NO: 47) mtqsvkvvekhvpiviekpsekeedtssedsieltvgkqpkpvtetrsld dleaimkagktklledhevvklslegklplyalekqlgdntravgirrsi isqqsntktletsklpylhydydrvfgaccenvigymplpvgvagpmnid gknyhipmattegclvastmrgckainagggvttvltqdgmtrgpcvsfp slkragaakiwldseeglksmrkafnstsrfarlqslhstlagnllfirf rtttgdamgmnmiskgvehslavmvkeygfpdmdivsvsgnyctdkkpaa inwiegrgksvvaeatipahivksvlksevdalvelnisknligsamags vggfnahaanlvtaiylatgqdpaqnvessncitlmsnvdgnllisvsmp sievgtigggtilepqgamlemlgvrgphietpganaqqlariiasgvla aelslcsalaaghlvqshmthnrsqaptpakqsqadlqrlqngsnicirs 1E. Production of pMB4692 (URA3 tef1p-crtZ) Encoding N. aromaticovans Carotene Hydroxylase.

The following carotene hydroxylase (CrtZ) ORF sequence was synthesized de novo based on protein sequence of Novosphingobium aromaticivorans, using Y. lipolytica codon bias:

(SEQ ID NO: 48) 5′-ttctagacacaaaaatgggtggagccatgcagaccctc gctgctatcctgatcgtcctcggtacagtgctcgctatggagtttgtcgc ttggtcttctcataagtatatcatgcatggcttcggatggggatggcata gagaccatcacgagccccatgagggatttcttgagaagaatgacttatac gccatcgttggcgctgccctctcgatactcatgtttgccctcggctctcc catgatcatgggcgctgacgcctggtggcccggaacctggatcggactcg gtgtcctcttctatggtgtcatctataccctcgtgcacgacggtctggtg caccaacgatggtttagatgggtgcctaaacgaggttacgccaaacgact cgtgcaggcccataagctgcaccacgccaccattggcaaggaaggaggcg tctcattcggtttcgtgttcgcccgagatcccgccgttctgaagcaggag cttcgagctcaacgagaagcaggtatcgccgtgctgcgagaggctgtgga cggctagacgcgt

This sequence was cleaved using XbaI and MluI and ligated, along with an Acc65I-NheI EF1 promoter fragment from pMB4629, to pMB4662 cut with Acc65I and MIA to produce pMB4692. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtZ protein of pMB4692 is as follows:

(SEQ ID NO: 49) mggamqtlaailivlgtvlamefvawsshkyimhgfgwgwhrdhhepheg flekindlyaivgaalsilmfalgspmimgadawwpgtwiglgvlfygvi ytlvhdglvhqrwfrwvpkirgyakirlvqahklhhatigkeggvsfgfv fardpavlkqelraqreagiavlreavdg 1F. Production of pMB4698 (ADE1 tef1p-crtW), Encoding Carotene Ketolase Derived from an Environmental Sample.

The following carotene ketolase (CrtW) ORF sequence was synthesized de novo, based on protein sequence of an environmental sequence isolated from the Sargasso Sea (Genbank accession AACY01034193.1):

(SEQ ID NO: 50) 5′-ttctagacacaaaaatgactcgatctatttcctggcct tccacctactggcacctccagccctcctgttcttcttgggtcgcaaacga attctctcctcaagcccgaaaaggtctcgtcctcgctggtctcattggtt ccgcttggctgcttactctcggacttggcttttcccttcccctccatcaa acgagctggcttctcatcggttgtctcgttctccttagatctttcctgca caccggactttttatcgttgcccatgacgctatgcacgcttctcttgttc ctgaccaccctggccttaaccgttggattggacgtgtctgtcttctcatg tatgctggactctcctacaaaagatgctgccgaaatcaccgtcgacacca ccaagcccctgaaacagttgaagaccctgactaccaacgatgcactaaca acaatatcctcgactggtacgttcactttatgggaaattacctcggatgg caacaattgcttaatctctcttgcgtttggctcgctctcaccttccgtgt ttctgactactctgctcaattcttccacctgctccttttctctgtccttc ctctcatcgtctcctcctgtcaactcttcctcgtgggaacctggctgcca caccgacgaggcgctactactcgacccggcgttaccactcgatccctgaa cttccaccctgctctttccttcgctgcttgctaccacttcggttaccacc gtgaacaccatgaatctccctctactccttggttccaacttcctaaactc cgagaaggttctctcatctaaacgcgt

This sequence was cleaved using XbaI and MluI and ligated to pMB4629 cut with NheI and MluI to produce pMB4698. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtW protein of pMB4698 is as follows:

(SEQ ID NO: 51) mtrsiswpstywhlqpscsswvanefspqarkglvlagligsawlltlgl gfslplhqtswlligclvllrsflhtglfivahdamhaslvpdhpglnrw igrvcllmyaglsykrccrnhrrhhqapetvedpdyqrctnnnildwyvh fmgnylgwqqllnlscvwlaltfrvsdysaqffhlllfsvlplivsscql flvgtwlphrrgattrpgvttrslnfhpalsfaacyhfgyhrehhespst pwfqlpklregsli

Mutant alleles of this protein (e.g. L200M, F238L/I/V, including combinations thereof) can also be constructed and tested.

1G. Production of pMB4741 (ADE1 tef-crtW), Encoding Aurantimonas Carotene Ketolase.

The following carotene ketolase (CrtW) ORF sequence was synthesized de novo based on protein sequence of Aurantimonas sp. S185-9A1, using Y. lipolytica codon bias:

(SEQ ID NO: 52) ctctagacacaaaaatgtcttcctttgcccctatgaatgatgttgctatt cctgccggtcaagctcctttctctgcctgtactagaaaacctgtcctgag accttttcaagctgccatcggtcttacactcgccggatgtgttatctctg cttggattgcaatccacgttggagctgtctttttcctcgatgtcggttgg cgaacccttcctgttgttcctgtcctcattgccgttcagtgctggctcac ggtcggtctttttattgtcgcacacgatgctatgcacggctccctcgctc ctggttggccacgacttaacgctcgaattggtgccttcatcctcaccatc tacgctggattcgcttggagacgtgtccgaggagctcacatggcccatca cgacgcccctggtactgccgatgaccctgacttctttgttgatgaacctg accgattttggccttggtttcgagctttcttccttagatattttggacgt cgatctattctctttgtttgcacagttgtcaccgtttacattctggtcct tggagcccctgttcttaatgttgttctcttttacggtcttccttcccttc tgtcttctcttcaactcttttactttggaacttttcgtcctcaccgtcat gaagaagatgatttcgttgacgcccataatgcccgatctaatgaatttgg ttacatcgcctccctcctttcttgctttcactttggataccatcacgaac atcatgccgagccgtgggtcccttggtggggtcttccttctcaatggcgc cagagacaagcctcttcttcccgacaggtcccgggcggccgagacgctgc tgacgccgctggagcatctcgacaacctgccggacgataccgatctgttt cttctcgaggtcgaaatcaggcccgttctcccgcttctggtcgaaacgaa caaatgagataaacgcgt

This sequence was cleaved using XbaI and MluI and ligated to pMB4629 cut with NheI and MluI to produce pMB4741. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtW protein of pMB4741 is as follows:

(SEQ ID NO: 53) mssfapmndvaipagqapfsactrkpvlrpfqaaigltlagcvisawia ihvgavffldvgwrtlpvvpvliavqcwltvglfivahdamhgslapgw prlnarigafiltiyagfawrrvrgahmahhdapgtaddpdffvdepdr fwpwfrafflryfgrrsilfvctvvtvyilvlgapvlnvvlfyglpsll sslqlfyfgtfrphrheeddfvdahnarsnefgyiasllscfhfgyhhe hhaepwvpwwglpsqwrqrqasssrqvpggrdaadaagasrqpagryrs vssrgrnqarspasgrneqmr

Mutant alleles of this protein (e.g. L201M, A232V/I/L, F240L/I/V, including combinations thereof) can also be constructed and tested.

1H. Production of pMB4735 (ADE1 tef-crtW), Encoding P. bermudensis Carotene Ketolase.

The following carotene ketolase (CrtW) ORF sequence was synthesized de novo based on protein sequence of Parvularcula bermudensis, using Y. lipolytica codon bias:

(SEQ ID NO: 54) ctctagacacaaaa atggaccctaccggagacgttactgctagccctcg acctcaaaccaccattcctgtccgacaagcactctggggacttagcctt gctggagccatcatcgccgcatgggtttttatgcacattggtttcgttt tttttgccccccttgatcctatcgttctcgccctcgccccagttattat tcttcttcaatcctggctttctgttggtctttttattatttctcacgac gcaattcacggttccctcgcccctggacgacccgcctttaatagagcca tgggacgactctgcatgacactttacgccggtttcgactttgaccgtat ggccgctgcacatcaccgacatcacagatcccctggaaccgccgctgac cccgatttttctgttgactcccctgatcgacctctcccttggtttggag ctttcttccgacgttactttggctggagaccttttcttaccgttaacgc tgtcgtctttacctactggcttgttcttggagctaaccctgttaatatt gttctcttttatggcgttcctgcactcctttccgccggacagctctttt actttggtacatttctccctcaccgacacgaacgacaaggctttgctga tcaccaccgagcacgatccgtccgatccccttacatgctttctcttgtt acttgttaccactttggaggctatcatcacgaacatcatctctttccac acgaaccctggtggcgcctgcctcaacgaggaggttgggaacgtgacag acgaaagagaaccggcccttaa cgcgt

This sequence was cleaved using XbaI and MluI and ligated to pMB4629 cut with NheI and MluI to produce pMB4735. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtW protein of pMB4735 is as follows:

(SEQ ID NO: 55) mdptgdvtasprpqttipvrqalwglslagaiiaawvfmhigfvffapl dpivlalapviillqswlsvglfiishdaihgslapgrpafnramgrlc mtlyagfdfdrmaaahhrhhrspgtaadpdfsvdspdrplpwfgaffrr yfgwrpfltvnavvftywlvlganpvnivlfygvpallsagqlfyfgtf lphrherqgfadhhrarsvrspymlslvtcyhfggyhhehhlfphepww rlpqrggwerdrrkrtgp

Mutant alleles of this protein (e.g. L190M, M110I/V/L, F229L/I/V, including combinations thereof) can also be constructed and tested.

1I. Production of pMB4778 (URA3 tef-crtZ), Encoding P. bermudensis Carotene Hydroxylase.

The following carotene hydroxylase (CrtZ) ORF sequence was synthesized de novo based on protein sequence of Parvularcula bermudensis, using Y. lipolytica codon bias:

(SEQ ID NO: 56) ctctagacacaaaa atgactctcgctctctggcaaaagatcaccctcgt ccttggttccgctgctctgatggaaggatttgcttggtgggcccataga tatattatgcacggttggggatgggcttggcatagagatcatcatgaac ctcacgacaaagtttttgaaaaaaatgacctgtttgctgtggtttttgg ctcgttcgcatttggtttgttcatcgtcggttacctttattggccacct gtttggtacgttgctgctggcatcactctttacggacttctttacgcat ttgttcatgacggtttggttcatcaacgttggccctggcatttcatgcc taaacgaggatacctccgaagactggttcaagctcacaaacttcatcat gctgttacaacacaaggcggaaatgtttcgtttggattcgtccttgccc ctgaccctagacatcttagagaaaaacttagacaatttcgtgctgaaag acatcgtgcccttgccgccgaaggtgcttcctcctctgaccctcgtgtt cccccttttcgaaaagttcaagacgtttaa acgcgt

This sequence was cleaved using XbaI and MluI and ligated to pMB4751 cut with NheI and MluI to produce pMB4778. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtZ protein of pMB4778 is as follows:

(SEQ ID NO: 57) mtlalwqkitlvlgsaalmegfawwahryimhgwgwawhrdhhephdkv fekndlfavvfgsfafglfivgylywppywyvaagitlygllyafvhdg lvhqrwpwhfmpkrgylrrlvqahklhhavttqggnvsfgfvlapdprh lreklrqfraerhralaaegasssdprvppfrkvqdv 1J. Production of pMB4719 (URA3 tef-crtZ), Encoding E. litoralis Carotene Hydroxylase.

The following carotene hydroxylase (CrtZ) ORF sequence was synthesized de novo based on protein sequence of Erythrobacter litoralis, using Y. lipolytica codon bias:

(SEQ ID NO: 58) ctctagacacaaaa atgagctggtgggctatcgctcttattgtctttgg tgctgtcgttggaatggaattttttgcttggttcgctcataagtacatt atgcatggttggggatggagctggcaccgagatcatcacgaacctcacg ataatactcttgaaaaaaacgaccttttcgccgttgtctttggctcggt tgccgcacttctgtttgttattggagctctctggtctgatcctctctgg tgggcagcagttggtattacattgtatggcgtcatttacactctggttc acgacggacttgttcatcaacgttactggcgttggacccctaagcgagg ttatgctaagagacttgtccaggcccatcgacttcatcacgctactgtt ggaaaggaaggaggtgtttcttttggttttgtgttcgcccgagatcctg ctaagttgaaagccgaattgaaacaacaaagagaacagggacttgccgt cgttcgagattctatgggagcataa acgcgt

This sequence was cleaved using XbaI and MluI and ligated to pMB4691 cut with NheI and MluI to produce pMB4719. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtZ protein of pMB4719 is as follows:

(SEQ ID NO: 59) mswwaialivfgavvgmeffawfahkyimhgwgwswhrdhhephdntle kindlfavvfgsvaallfvigalwsdplwwaavgitlygviytlvhdgl vhqrywrwtpkrgyakrlvqahrlhhatvgkeggvsfgfvfardpaklk aelkqqreqglavvrdsmga 1K. Production of pMB4812, Encoding N. crassa Phytoene Synthase/Lycopene Cyclase, al-2.

Exon 1 of al-2 was synthesized by annealing the following oligonucleotides:

MO5017: (SEQ ID NO: 60) 5′-CTAGACACAAAAATGTACGACTACGCCTTCGT; MO5018: (SEQ ID NO: 61) 5′-GCACCTGAAGTTCACCGTGCCCGCGGTTCCAA; MO5019: (SEQ ID NO: 62) 5′-GTGCACGAAGGCGTAGTCGTACATTTTTGTGT; MO5020: (SEQ ID NO: 63) 5′-CGCGTTGGAACCGCGGGCACGGTGAACTTCAG, and ligating them to pMB4603 that had been cleaved with NheI and MluI, to create pMB4811. Exon2 was amplified from N. crassa (Fungal Genetic Stock Center #3200) genomic DNA, using MO5016 (5′-CCCGCGGCGGTACTTCT (SEQ ID NO:64)) and MO5013 (5′-CCGTCTCTACAGCAGGATCAGGTCAATGC (SEQ ID NO:65)), and inserted into pCR-TOPO (Invitrogen), to create pMB4809. Exon 3 was similarly amplified with MO5014 (5′-CCGTCTCACTGTACTCCTTCTGTCGCCTG (SEQ ID NO:66)) and MO5015 (5′-CACGCGTCTACTGCTCATACAACGCCCT (SEQ ID NO:67)), and cloned into the same vector to create pMB4810. The 0.9 kb SacII-BsmBI fragment from pMB4809 was ligated together with the 0.9 kb BsmBI-MluI fragment from pMB4810 into SacII-MluI-cleaved pMB4811, to create pMB4812, which expresses al-2 from the TEF1 promoter. The resulting nucleic acid coding sequence and encoded al-2 protein of pMB4812 are as follows:

(SEQ ID NO: 68) atgtacgactacgccttcgtgcacctgaagttcaccgtgcccgcggcgg tacttctcaccgctatcgcctaccccattctcaacaggatacatctcat ccaaacaggcttcctcgtcgtcgtcgcctttaccgccgctctgccatgg gatgcctacttgattaagcacaaagtatggtcttacccaccagaagcca ttgttgggccgcgtttgcttggaattccctttgaagagctgttcttctt tgtgatacagacttacatcacggcgctcgtatacatcctcttcaacaag ccggtgctgcacgcgttgcacctcaacaatcaacaaaacccgccagcat ggatgagggttgtcaaggttaccggccaggtagtcctcgtagccttgtc ggtatggggatggaatgccgctcaggttcatcaggaaacaagctatctc ggcttgatccttgtttgggcttgtccgttcttactggctatctggaccc tcgctgggcgcttcattctcagcctaccctggtacgcgacggtgctccc gatgttcctacccaccttctatctttgggcggtagacgagtttgccttg cacaggggtacttggtccatcggatcggggacgaagctcgatttttgtc tgtttggcaagttggacattgaagaagccacgttcttcctggtgaccaa catgctcatcgttggcggtatggccgcgttcgatcaatatctggccgtc atttacgctttcccaactctgttccccaaggtcaaccggtatccgacaa ctcatatgcttcttcaaagccgtcttatcaacacttccaggtacgatct tgagcgcattgagggcctgagagaagcggtcgagagactgcgcctgaag agcaggagtttttacctggccaattcgctcttttctggtcgactccgca ttgacctgatcctgctgtactccttctgtcgcctggctgatgatctagt cgacgacgccaaatctcgccgtgaggtcttgtcctggaccgcgaagctg aaccacttccttgatctgcactacaaggacgcggacgccaccgaggacc ccaagaaaaaggcggagcgaatcgacgcctacatcaagacagcgttccc tccctgtgcctaccaagccctccacctcctgcccactcacattcttcct cccaagcctctttacgatctcatcaagggtttcgagatggactctcaat tcaccttccacggtacttccgactctacggatctccaataccccatcgc cgacgacaaggaccttgagaactacgctatctatgtcgccggtaccgtc ggcgagctctgcatcgccctcatcatctaccactgcctgccagacatgt cggacactcagaagcgcgagctcgagaccgccgcgtgccggatgggcat cgcgctgcagtacgtcaacatcgctcgtgacatcgtcgtcgacgcacgt atcgggcgcgtttacttgcctaccacctggctcaagaaggaagggttga cgcacaagatggtcttggagaaccccgagggtcccgaggtcattgagcg gatgagaagacggcttttggaaaatgcgtttgagctgtatgggggcgcg aggcctgagatgcaacggataccgagcgaggctaggggcccgatgattg gtgccgttgaaaattacatggcgattggaagggtgttgagggagaggaa ggaggggacggtgtttgtgaggatggaggggagggctacggtcccgaag cgaaggaggttgagcacgctgttgagggcgttgtatgagcagtag; (SEQ ID NO: 69) mydyafvhlkftvpaavlltaiaypilnrihliqtgflvvvaftaalpw daylikhkvwsyppeaivgprllgipfeelfffviqtyitalvyilfnk pvlhalhlnnqqnppawmrvvkvtgqvvlvalsvwgwnaaqvhqetsyl glilvwacpfllaiwtlagrfilslpwyatvlpmflptfylwavdefal hrgtwsigsgtkldfclfgkldieeatfflvtnmlivggmaafdqylav iyafptlfpkvnryptthmllqsrlintsrydlerieglreaverlrlk srsfylanslfsgrlridlillysfcrladdlvddaksrrevlswtakl nhfldlhykdadatedpkkkaeridayiktafppcayqalhllpthilp pkplydlikgfemdsqftfhgtsdstdlqypiaddkdlenyaiyvagtv gelcialiiyhclpdmsdtqkreletaacrmgialqyvniardivvdar igrvylpttwlkkeglthkmvlenpegpeviermrrrllenafelygga rpemqripseargpmigavenymaigrvlrerkegtvfvrmegratvpk rrrlstllralyeq 1L. Production of pMB4846 (URA3 tef-crtZ), Encoding an Erythrobacter Carotene Hydroxylase.

The following carotene hydroxylase (CrtZ) ORF sequence was synthesized de novo based on protein sequence of Erythrobacter sp. NAP1, using Y. lipolytica codon bias:

(SEQ ID NO: 70) ctctagacacaaaaatgtcttggcctgccgctattgcagttacacttgg tgcccttatttttatggaattctttgcttggtacgctcacaaatacatt atgcatggatggggatggggttggcacagagaccatcacgaacctcacg acaacaaactggaaaaaaatgacctgttcgctgtggttttcggaacaat taacgctggtatgtatatttttggtgctctttattgggatgctttgtgg tgggctgcacttggagttaatctttacggagtgatttacgcccttgttc atgacggactggttcatcaaagatttggaagatacgtccctaaaaacgc atacgctaaacgacttgttcaagcacacagattgcatcacgctactatc ggtaaagaaggaggagtgtccttcggattcgttcttgctcgagaccctg ctaaacttaaagccgaacttaaacgacaatctcaatccggagaagctat tgttcgagaatccgccggagcctaaacgcgt

This sequence was cleaved using XbaI and MluI and ligated to pMB4691 cut with NheI and MluI to produce pMB4846. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtZ protein of pMB4846 is as follows:

(SEQ ID NO: 71) mswpaaiavtlgalifmeffawyahkyimhgwgwgwhrdhhephdnkle kndlfavvfgtinagmyifgalywdalwwaalgvnlygviyalvhdglv hqrfgryvpknayakrlvqahrlhhatigkeggvsfgfvlardpaklka elkrqsqsgeaivresaga 1M. Production of pMB4835 (URA3 tef-crtZ), Encoding an S. alaskensis Carotene Hydroxylase.

The following carotene hydroxylase (CrtZ) ORF sequence was synthesized de novo based on protein sequence of Sphingopyxis alaskensis, using Y. lipolytica codon bias:

(SEQ ID NO: 72) ctctagacacaaaaatgagccaccgaagagatccaggacttagaagaga cgacgcacgatctatggcctcctgtctcagacgagcttacaacccccac atgtccctgcctgcaattttgtttttggttcttgctactgtcattgcaa tggaaggagtcgcctgggcatcccacaaatacatcatgcacggatttgg atgggcctggcacagagaccaccatgaaccccacgacaatcgactcgag aaaaacgacctgtttgccctgttcggagccgctatgtctatttctgcct tcgctattggttctcctatgattatgggtgcagctgcctggaagcctgg aacttggattggacttggtattcttctttacggtattatctacacactc gttcacgacggccttgtgcaccaaagatactttcgatgggtcccacgac gaggttacgcaaaacgacttgttcaagcacacaaacttcatcacgctac aatcggaaaagagggaggagtttctttcggatttgtttttgctcgtgac cctgctaaacttaaagccgaactgaaagcacaacgagaagctggtattg cagtcgtcagagaagcccttgctgactaaacgcgt

This sequence was cleaved using XbaI and MluI and ligated to pMB4691 cut with NheI and MluI to produce pMB4835. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtZ protein of pMB4835 is as follows:

(SEQ ID NO: 73) mshrrdpglrrddarsmasclrraynphmslpailflvlatviamegva washkyimhgfgwawlirdhhephdnrlekndlfalfgaamsisafaig spmimgaaawkpgtwiglgillygiiytlvhdglvhqryfrwvprrgya krlvqahklhhatigkeggvsfgfvfardpaklkaelkaqreagiavvr ealad 1N. Production of pMB4845 (URA3 tef-crtZ), Encoding an R. biformata Carotene Hydroxylase.

The following carotene hydroxylase (CrtZ) ORF sequence was amplified from genomic DNA extracted from Robiginitalea biformata:

(SEQ ID NO: 74) cacaatctagacacaaaa atgacagtcttgatttggatcgcaattttcc tggccaccttctgcttcatggaattcatggcctggtttacgcataaata tatcatgcacggtttcctctggagccttcataaggaccaccataaaaag gaccacgacagttggtttgagcgaaacgacgccttctttctattttatg cgatagtctccatgtcctttatcggggccgccgtgaacacgggattctg gcaggggtggcccatcggcctgggcatcctcgcttacgggattgcctac tttatcgtacacgatatctttatccatcagcggttcaagctctttcgca atgcgaataactggtacgcgcggggtatccgcagggcccataaaatcca ccacaagcacctgggaaaagaggaaggggaatgcttcgggatgctgttt gtcccatttaagtacttccggaagacctga acgcgtttgtg

This sequence was phosphorylated and ligated to pBluescriptSK⁻ that had been cleaved with EcoRV and dephosphorylated, to create pMB4824. The XbaI-MluI fragment from pMB4824 that contains crtZ was ligated to pMB4691 cut with NheI and MluI to produce pMB4845. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtZ protein of pMB4845 is as follows:

(SEQ ID NO: 75) mtvliwiaiflatfcfmefmawfthkyimhgflwslhkdhhkkdhdswf erndafflfyaivsmsfigaavntgfwqgwpiglgilaygiayfivhdi fihqrfklfrnannwyargirrahkihhkhlgkeegecfgmlfvpfkyf rkt 1O. Production of pMB4837 (URA3 tef-crtZ), Encoding an X. autotrophicus Carotene Hydroxylase.

The following carotene hydroxylase (CrtZ) ORF sequence was amplified from genomic DNA extracted from Xanthobacter autotrophicus:

(SEQ ID NO: 76) cacaatctagacacaaaa atgtccaccagcctcgccttcctcgtcaacg cgctcatcgtgatcgccacggtcgccgccatggaaggggtggcctggg ccgcgcacaaatatgtcatgcacggcttcggctggggctggcacaagtc ccaccacgagccgcgcgagggcgtgttcgagcgcaacgacctttatgc gctgctgttcgcaggcatcgccatcgccctcatctacgcgttccgcaat ggcggcgcgctgctgtgggtgggcgtggggatgacggtctacggcttcc tttatttcttcgtgcacgacggcatcacccaccagcgctggccgttcc gctacgtgccgcgcaacggctatctcaagcgcctggtgcaggcccacc ggctgcaccatgcggtggatggcaaggagggctgcgtctccttcggctt catctatgccccgccgcctgccgacctgaaggccaagctgaagaagct gcacggcggcagcctgaagcagaacgaggcggcggaatag acgcgt ttgtg

This sequence was phosphorylated and ligated to pBluescriptSK⁻ that had been cleaved with EcoRV and dephosphorylated, to create pMB4823. The XbaI-HindIII (filled in with Klenow) fragment from pMB4823 that contains crtZ was ligated to pMB4691 cut with NheI and MluI (filled in with Klenow) to produce pMB4837. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtZ protein of pMB4837 is as follows:

(SEQ ID NO: 77) mstslaflvnaliviatvaamegvawaahkyvmhgfgwgwhkshhepr egvferndlyallfagiaialiyafrnggallwvgvgmtvygflyffvh dgithqrwpfryvprngylkrlvqahrlhhavdgkegcvsfgfiyappp adlkaklkklhggslkqneaae 1P. Production of pMB4850 (URA3 tef-crtZ), Encoding a P. putida Carotene Hydroxylase.

The following carotene hydroxylase (CrtZ) ORF sequence was amplified from genomic DNA extracted from Pseudomonas putida (this sequence encodes a valine rather than a leucine at the second position, due to N-end rule considerations):

(SEQ ID NO: 78) tctctctagacacaaaa atggtgttcaatctcgccatattgttcggcac cctggtggccatggagggcgttggtacgctggctcacaagtacatcatg catggctggggctggtggctgcaccgatcgcaccatgagccacacctg ggcatgctcgaaaccaacgacctgtacctggtggccctggggctgatc gccacggcgctggtggcgctgggcaaaagtggttatgcgcctttgcagt gggtgggcggtggtgtggcaggctatggagcactgtatgtactggccca cgacggtttctttcaccggcactggccgcgcaagccgcggccggtcaa ccgctacctgaaacgcttgcaccgcgcgcaccgcttgcaccatgcggt gaaggggcgcacggggagcgtgtcgttcgggttcttctatgcgccgccg ctgaaggtgttgaagcagcaattgcgcagcaggcgcagccaatcgtga a cgcgtgagacgttgtg

This sequence was phosphorylated and ligated to pBluescriptSK⁻ that had been cleaved with EcoRV and dephosphorylated, to create pMB4847. The XbaI-MluI fragment from pMB4847 that contains crtZ was ligated to pMB4691 cut with NheI and MluI to produce pMB4850. The nucleic acid coding sequence is depicted in bold underline above. The resulting encoded CrtZ protein of pMB4850 is as follows:

(SEQ ID NO: 79) mvfnlailfgtlvamegvgtlahkyimhgwgwwlhrshhephlgmletn dlylvalgliatalvalgksgyaplqwvgggvagygalyvlahdgffhr hwprkprpvnrylkrlhrahrlhhavkgrtgsvsfgffyapplkvlkqq lrsrrsqs

Example 2 Engineering Yarrowia lipolytica for Increased Carotenoid Production

2A. Production of Y. Lipolytica Expressing Geranylgeranylpyrophosphate Synthase and Phytoene Dehydrogenase:

MF350 (MATB ura2-21 leu2-35 ade1) was transformed with pMB4591 (tef1p-GGS1) that had been cleaved upstream of URA5 with SspI; a Ura⁻ transformant carrying the plasmid at the ura2 locus was identified and named MF364. It was subsequently transformed with pMB4638 (tef1p-carB) that had been cleaved at ADE1 with SspI and a prototrophic transformant was chosen that harbored the plasmid at the ade1 locus. This strain was named MF502.

2B. Production of Y. Lipolytica Expressing Geranylgeranylpyrophosphate Synthase, Phytoene Dehydrogenase and Phytoene Synthase/Lycopene Cyclase

MF502 was transformed with pMB4705 (tef1p-carRP[i⁻]) that had been treated with SspI. Ninety percent of the prototrophic transformants were very orange on YPD agar plates, and one, MF719, produced greater than 10 mg carotene per g dry cell weight (DCW) after four days of growth in YPD at 30° C.

2C. Production of Y. Lipolytica Expressing Phytoene Synthase/Lycopene Cyclase and Phytoene Dehydrogenase:

ATCC201249 (HATA ura3-302 leu2-270 lys8-11) was transformed with SspI-cleaved pMB4628. Hundreds of Leu⁺ colonies were pooled, re-grown, and transformed with pMB4660 (tef1p-carB) that had been cleaved upstream of URA3 with SalI. One colony that was noticeably yellow after 5 days at 30° C. on YNBglut media (per liter: 1.7 g yeast nitrogen base, 1 g monosodium glutamate, 1% glucose) plus 0.6 mM lysine was selected, named MF447, and found to produce 0.2 mg carotene per gram dry cell weight after 4 days of growth in YPD.

MF447 was challenged with 1 g/L 5-fluoroorotic acid and Ura⁻ segregants selected. Surprisingly, they were all found to retain the identical yellow appearance of their parent, implying that the loss of a functional URA3 gene did not coincide with the loss of a functional CarB enzyme. Southern analysis demonstrates that two fragments from a KpnI-HindIII digest of MF447 DNA contain URA3p-hybridizing sequences, only one of which also hybridizes to carB. The other is absent in MF578, the Ura3⁻ segregant chosen for further manipulation. Plasmid rescue and analysis of the DNA sequence surrounding tef-carB in MF578 confirmed the absence of nearby URA3 sequences. Plasmid rescue and analysis of the DNA sequence encompassing the carRP intron in MF447 revealed that exons 1 and 2 were contiguous and were each separated by an intron sequence that lacked the original internal SspI site (present in pMB4628). The sequence of this region shows a seven-base pair deletion (AATATTA) that would restore the proper frame to an unspliced message. Partial intron sequence comprising the sequence where the deletion occurred is shown as follows:

(SEQ ID NO: 80) ACAAACAAATGATGTGCCGCATCGCATTTTAATATTAACCATTGCATACA CAG.

Predicted partial amino acid sequence comprising this intron, if unspliced, is as follows:

(SEQ ID NO: 219) KAW VSKQTNDVPHRILIPLHTQ HLA . . . (VSKQTNDVPHRILIPLHTQ (SEQ ID NO: 81) is intron encoded) 2D. Production of Y. Lipolytica Expressing Phytoene Synthase/Lycopene Cyclase, Phytoene Dehydrogenase and Geranylgeranylpyrophosphate Synthase:

MF578 was transformed with pMB4683 (tef1p-GGS1) that had been cleaved with SalI (upstream of URA3) or with StuI (within the GGS1 ORF). Ura⁺ Leu⁺ colonies in both cases appeared bright orange on YNBglut+Lysine and on YPD, and several produced greater than 4 mg carotene per gram of dry cell weight when grown as above. One, MF633, contained a single copy of the plasmid at the GGS1 locus, as inferred from Southern analysis. The others arose by non-homologous or more complex integrations.

2E. Production of Y. Lipolytica Expressing Phytoene Synthase/Lycopene Cyclase, Phytoene Dehydrogenase and Geranylgeranylpyrophosphate Synthase:

MF364 was crossed with MF578, and spores from the resulting diploid were plated on YPD for two to three days at 30° C. White Leu⁻ Ade⁻ Ura⁻ colonies were screened for the presence of tefp-carB and tefp-GGS1 and for the absence of tefp-carRP by PCR. Thirteen colonies meeting these criteria, as well as displaying resistance to 5-fluorootic acid, an indication that they harbor the ura3-302 allele, were chosen as hosts for further modifications.

One such strain, MF731, was transformed with pMB4705 cut with BbvCI, and one Leu⁺ orange colony, MF740, produced 6 mg of β-carotene per g DCW after four days of growth in YPD at 30° C.

Another tefp-carB tefp-GGS1 strain from the same cross, MF739, was transformed with pMB4705 cut with BbvCI, and one Leu⁺ orange colony, MF746, produced 8 mg of β-carotene per g DCW after four days of growth in YPD at 30° C. When this strain was transformed with pMB4812 (expressing N. crassa al-2 protein) treated with SspI, the Leu⁺ transformants were less orange than parallel pMB4705 Leu⁺ transformants, and after 4 days of growth in YPD, produced about half the amount of β-carotene as pMB4705 transformants. In addition, the pMB4812 transformants produced significant amounts of γ-carotene (˜40% of total carotene.).

2F. Expression of a Truncated Form of HMG-CoA Reductase Results in Increased Carotenoid Production in Y. Lipolytica Expressing Phytoene Synthase/Lycopene Cyclase, Phytoene Dehydrogenase, and Geranylgeranylpyrophosphate Synthase:

In order to increase carotenoid production, carbon flow through the isoprenoid pathway is enhanced by introducing a truncated variant of the HMG-CoA reductase gene.

MF740 was transformed with pMB4637 treated with SnaBI, and Ade⁺ colonies were selected. One such colony, MF760, was shown to produce about 20 mg β-carotene per g DCW after four days of growth in YPD at 30° C. This strain was also the subject of several fermentor studies outlined in Example 5. In addition, MF740 was also transformed with MB4714 treated with AflII and Ura⁺ colonies, were selected. One such colony was designated MF779 (see Example 2G). MF746 was also transformed with pMB4637 treated with SnaBI, and Ade⁺ colonies were selected. One such colony, MF946, was shown to produce greater than 35 mg β-carotene per g DCW after four days of growth in YPD at 30° C.

MF760 was also transformed with pMB4691 (empty vector) cut with SalI, creating the prototroph MF858.

2G. Production of Y. Lipolytica Expressing Carotene Ketolase, a Truncated Form of HMG-CoA Reductase, Phytoene Synthase/Lycopene Cyclase, Phytoene Dehydrogenase and Geranylgeranylpyrophosphate Synthase:

MF779 was transformed with either pMB4735 or pMB4741 cleaved with SnaBI., and a red prototrophic colony was chosen from each transformation: MF838 (pMB4735) and MF840 (pMB4741). After 4 days of growth in YPD, MF838 produced 25 mg canthaxanthin per g DCW, and MF840 produced 14 mg canthaxanthin and 30 mg echinenone per g DCW. Only trace levels of β-carotene were produced. These strains are the subject of fermentor studies described in Example 5.

In addition, MF740 was transformed with pMB4735 cleaved with SnaBI, and a red Ade⁺ colony was chosen for further manipulation and designated MF889 (See example 2I).

2H. Manipulation of the Y. Lipolytica ERG9 Gene to Enhance Carotenoid Production.

In order to decrease the expression of Erg9 (squalene synthase) in a carotenoid-producing yeast, pMB4789, containing the following cassette, was constructed using standard molecular techniques. The 4.8 kb fragment contains the Y. lipolytica URA3 gene flanked by the ERG9 ORF and the ERG9 terminator.

Thus this fragment comprises the sequence: GATCtcgttctgctcgggtagatc (SEQ ID NO:82)-ERG9 (promoter and ORF)-gtgctctgcggtaagatcgACTAGTggtgtgttctgtggagcattc (SEQ ID NO:83) URA3 (promoter, ORF, and terminator) ccaccactgcactaccactacacCTCGAGCATGCATcaggaaggactctccctgtggt (SEQ ID NO:84)-ERG9terminator-gtgttatggctctacgtgaagGGGCCC (SEQ ID NO:85). (Capital letters: restriction sites [engineered for assembly]) In addition, it was found that a mutation was generated during cloning that changed the coding sequence of ERG9 as follows: (cccgacgttAtccagaagaac (SEQ ID NO:86); F317I in the encoded protein).

Two overlapping fragments from this cassette, a 2.4 kb AlwNI-SmaI fragment and a 1.9 kb AlwNI-AflII fragment, were cotransformed into MF760 and Ura⁺ colonies were selected. PCR analysis showed that one, designated MF921, contains the erg9::URA3 cassette replacing the wild type ERG9 gene. MF921 produced greater than 30 mg β-carotene per g DCW after 4 days of growth at 30° C. in YPD.

2I. Production of Y. Lipolytica Expressing Carotene Hydroxylase, Phytoene Synthase/Lycopene Cyclase, Phytoene Dehydrogenase and Geranylgeranylpyrophosphate Synthase:

MF740 was transformed with pMB4837 cleaved with SalI, and a Ura⁺ colony was selected and designated MF1011. MF1011 produced 6 mg of zeaxanthin and 1.5 mg of β-carotene per g DCW after 4 days of growth at 30° C. in YPD.

2J. Production of Y. Lipolytica Expressing Carotene Hydroxylase, Carotene Ketolase, Phytoene Synthase/Lycopene Cyclase, Phytoene Dehydrogenase and Geranylgeranylpyrophosphate Synthase:

MF889 was transformed with pMB4837 cleaved with SalI, and a prototrophic colony was selected and designated MF1016. MF1016 produced 1.5 mg of astaxanthin and 3 mg of canthaxanthin per g DCW after 4 days of growth at 30° C. in YPD.

Example 3 Extraction of Carotenoids

3a: Total Extraction of Carotenoids from Yarrowia lipolytica Cells

Yarrowia lipolytica cultures to be tested for carotenoid production were grown in 20 ml YPD medium (1% yeast extract, 2% peptone, 2% glucose) in 125 flasks at 30° C. Following incubation for 72-96 hr, the cultures were harvested by centrifugation and the solvent extractions were performed to determine carotenoid form and quantity. Dry cell weights were determined by transferring 1.8 ml of each culture to an Eppendorf tube, which was then centrifuged to pellet the cells, and then the pellet washed twice with 1 ml water. After the second wash, the cells were resuspended in water and transferred to a pre-weighed snap-cap tube with a hole poked in the top, frozen, and then lyophilized overnight. After drying to constant weight, the tube was weighed in order to calculate dry cell weight (mg dry cell weight/ml).

The carotenoid content of the culture was calculated by solvent extraction from 0.25 ml of culture from the same shake flask culture. This 0.25 ml culture sample was transferred to a 2 ml screw-cap tube, the cells pelleted, and the supernatant aspirated. Such pelleted cells may be extracted immediately or frozen at −80° C. and stored.

An equal volume of cubic zirconia beads was added to cell pellets, along with 1 ml ice-cold extraction solvent (a 50/50 v/v mix of hexane and ethyl acetate containing 0.01% butylhydroxytoluene (BHT)). The mixture was then agitated (Mini-BeadBeater-8, BioSpec Products, Inc.) at maximum speed for 5 minutes at 4° C. The mixture was then spun at maximum speed for 1 minute, and the supernatant was collected and deposited in a cold 16 ml glass vial.

The remaining cell debris was re-extracted at least three times, without the addition of zirconia beads; all supernatants were pooled in the 16 ml glass vial. Following extraction, the glass vial was spun for 5 minutes at 2000 rpm at 4° C. in a Sorvall tabletop centrifuge, and the supernatant was transferred to a new cold 16 ml glass vial. A Speed Vac was used to concentrate the supernatant (room temperature in dark), and the samples were stored at −20° C. or −80° C. until immediately before HPLC analysis. Prior to HPLC analysis, the samples were resuspended in 1 ml ice-cold solvent and then transferred to a cold amber vial. Throughout the protocol, care was taken to avoid contact with oxygen, light, heat, and acids.

The use of a hexane:ethyl acetate (50:50) mixture to extract carotenoids efficiently extracted all carotenoids analyzed from Yarrowia even though the carotenoids possessed different polarity levels. For instance, in a strain containing β-carotene, γ-carotene, echinenone, and canthaxanthin, a hexane:ethyl acetate (50:50) mixture efficiently extracted all carotenoids even though echinenone and canthaxanthin, respectively, are progressively more polar than either β-carotene or γ-carotene. Given the high efficiency of extraction observed for all carotenoids with 50:50 hexane:ethyl acetate, these conditions were chosen as a “100%” standard against which the extraction efficiency of other conditions could be compared.

3B. Extraction of β-Carotene from Y. lipolytica MF858.

Y. lipolytica strain MF858 was grown as described in Example 3a and found to contain β-carotene as the dominant carotenoid. Extraction and breakage with hexane yielded an equal amount of β-carotene as was observed with a 50:50 hexane:ethyl acetate mixture.

3C. Extraction of Mixed Carotenoids from Y. Lipolytica MF838.

Y. lipolytica strain MF838 (example 2g) had previously been found to contain the following types carotenoids when extracted as described in Example 3a: β-carotene, γ-carotene, echinenone, and canthaxanthin. Extraction with 750 μl of hexane resulted in the following extraction efficiencies for each of the carotenoids (extraction efficiency is reported independently for each of the carotenoid species based on the total amount found by hexane:ethyl acetate extraction): β-carotene (79.3%), γ-carotene (82.4%), echinenone (42.6%), and canthaxanthin (8.0%).

When an identical aliquot of MF838 was extracted with 1 ml of ethanol (95%), the extraction efficiency of the same four carotenoids was as follows: β-carotene (53.6%), γ-carotene (71.3%), echinenone (39.9%), and canthaxanthin (28.0%). Thus ethanol can be used to extract both polar and nonpolar carotenoids from fungi (e.g. Y. lipolytica).

Example 4 Quantification of Carotenoid Production by HPLC

For carotenoid analysis, samples were resuspended in ice-cold extraction solvent (a 50/50 v/v mix of hexane and ethyl acetate containing 0.01% butylhydroxytoluene (BHT)). An Alliance 2795 HPLC (Waters) equipped with a Waters XBridge C18 column (3.5 μm, 2.1×50 mm) and Thermo Basic 8 guard column (2.1×10 mm) was used to resolve carotenoid at 25° C.; authentic carotenoid samples were used as standards. The mobile phases and flow rates are shown below (Solvent A=Ethyl Acetate; Solvent B=Water; Solvent C=Methanol; Solvent D=Acetonitrile). The injection volume was 10 μL. The detector is a Waters 996 photodiode array detector. The retention times for lipophilic molecules include astaxanthin (1.159 min), zeaxanthin (1.335), β-apo-8′-carotenal (2.86 min), ergosterol (3.11 min), lycopene (3.69 min), β-carotene (4.02 min), Canthaxanthin (2.50 min), Echinenone (3.38 min), and phytoene (4.13 min) Astaxanthin, zeaxanthin, β-apo-8′-carotenal, lycopene, β-carotene, canthaxanthin, and echinenone are detected at 475 nm, whereas ergosterol and phytoene were detected at 286 nm.

TABLE 28 Retention Times for Lipophilic Molecules Time Flow (min) (mL/min) % A % B % C % D Curve 0.50 0.0 20.0 0.0 80.0 3.00 1.00 20.0 0.0 0.0 80.0 6 4.50 1.00 80.0 0.0 20.0 0.0 6 5.00 1.00 0.0 0.0 100.0 0.0 6 6.00 1.00 0.0 0.0 100.0 0.0 6 6.50 1.00 0.0 20.0 0.0 80.0 6 7.00 0.50 0.0 20 0.0 80.0 6

Example 5A 2 Liter Fed-Batch Fermentation of β-Carotene Producing Strain MF760

FIG. 9A depicts the production and intracellular accumulation of phytoene and β-carotene by strain MF760 (Example 2F) when grown in fed-batch fermentation on various carbon sources. Fermentation medium and process parameters are described below. Carbon sources used were glucose, glycerol, or olive oil. Feeding was initiated during the early exponential growth phase at a rate of 15.2 ml/hr. This feed rate either continued until feed exhaustion or, when the dissolved oxygen (dO2) level of the culture reached 20% saturation, feed was added to maintain the dO2 at 20% (DO controlled feed). As seen in FIG. 9a , β-carotene accumulates in all fermentations from 1.5 to 2.2% of DCW weight. Substantial phytoene accumulation was observed in the constant feed fermentations but not in the DO controlled feed fermentations.

FIG. 9B depicts dry cell weight accumulation during the course of the fermentations. For each carbon source examined, constant feeding resulted in greater biomass production relative to the DO controlled feed fermentation. This was especially true for the olive oil fed-batch fermentations where the constant feed fermentation reached greater than 150 g/L DCW. This was expected as Y. lipolytica has been reported to accumulate greater than 40% its biomass as lipid when grown on oils under conditions of excess carbon and oxygen limitation (Pananikolaou et al., Appl. Microbiol. Biotechnol. 58:p 308, 2002) and was independent of nitrogen concentration.

Batch Medium—1 L

Carbon Source (one of the following): Glucose 60 g Glycerol 75 g Olive Oil 50 ml Yeast Nitrogen Base 4.1 g w/o Amino Acids and (NH₄)₂SO₄ (NH₄)₂SO₄ 6 g Uracil 72 mg Antifoam 204 (Sigma catalog A6426) 5 ml Feed Medium—1 L

Carbon Source (one of the following): Glucose 500 g Glycerol 500 g Olive Oil 500 ml (NH₄)₂SO₄ 72 g KH₂PO₄ 13.5 g MgSO₄ 5 g Inositol 70 mg Thiamine 10 mg Uracil 900 mg Trace Metal Solution 40 ml FeCl₃•6H₂O 2.7 g/L ZnCl₂•4H₂O 2.0 g/L CaCl₂•2H₂O 2.0 g/L Na₂MoO₄•2H₂O 2.0 g/L CuSO₄•5H₂O 1.9 g/L H₃BO₃ 0.5 g/L MnSO_(4•)H₂O 2.23 g/L Concentrated HCl 10 ml/L Vitamins Solution 40 ml Pantothenic acid 5.4 g/L Pyridoxine 1.4 g/L Niacin 6.1 g/L Folic acid 0.04 g/L Biotin 0.06 g/L Fermentation Parameter: pH 5.5, controlled Temp - 30° C. Air Flow - 1.4 lpm Agitation - 1200 rpm Inoculum - 200 ml overnight culture grown in YEP + 5% glucose

Example 5B 2 Liter Fed-Batch of β-Carotene Producing Strain MF760

FIG. 9c depicts the production and intracellular accumulation of β-carotene by strain MF760 when grown in fed-batch fermentation. In this fermentation, additions of olive oil were combined with a glucose feeding protocol. Medium and process parameters are described below. Both glucose and olive oil were present in the batch medium. Feeding of the glucose containing feed medium was initiated during the early exponential growth phase at a rate of 15.2 ml/hr, this feed rate continued until feed exhaustion. 25 ml of olive oil was added at 24, 48, and 72 hr.

As shown in FIG. 9c , this combined glucose and oil feeding protocol resulted in substantially higher DCW production when compared to glucose as the sole carbon source (example 5a). In addition, β-carotene accumulated to over 5% of the DCW at the end of the fermentation, higher then either the glucose or oil fermentations of example 5a.

Batch Medium—1 L

Glucose 40 g Olive Oil 50 ml Yeast Nitrogen Base 4.1 g w/o Amino Acids and (NH₄)₂SO₄ (NH₄)₂SO₄ 6 g Uracil 72 mg Antifoam 204 5 ml Feed Medium—1 L

Glucose 500 g (NH₄)₂SO₄ 72 g KH₂PO₄ 13.5 g MgSO₄ 5 g Inositol 70 mg Thiamine 10 mg Uracil 900 mg Trace Metal Solution 40 ml Vitamins Solution 40 ml Olive Oil Additions - 25 ml at 24, 48, and 72 hr Fermentation Parameters: pH 5.5, controlled Temp - 30° C. Air Flow - 1.4 lpm Agitation - 1150 rpm Inoculum - 200 ml overnight culture grown in YEP + 5% glucose + 2.5% olive oil

Example 5C 2 Liter Fed-Batch of Canthaxanthin Producing Strain MF840

FIG. 9d depicts the production and intracellular accumulation of canthaxanthin, echinenone and β-carotene by strain 840 (Example 2g) when grown in fed-batch fermentation. Medium and process parameters are described below. Both glucose and olive oil were present in the batch medium. Feeding of the glucose containing feed medium was initiated during the early exponential growth phase at a rate of 15.2 ml/hr; this feed rate continued until the dissolved oxygen reached 20%, at which time feed was added to maintain the dO2 at 20% (DO controlled feed) for the remainder of the fermentation.

As seen in FIG. 9d , the combined total amount of canthaxanthin, echinenone and β-carotene represented over 8% of the DCW at the end of the fermentation and demonstrates the ability of genetically engineered Y. lipolytica to produce and accumulate significant amounts of carotenoids.

Batch Medium—1 L

Glucose 40 g Olive Oil 50 ml Yeast Nitrogen Base 4.1 g w/o Amino Acids and (NH₄)₂SO₄ (NH₄)₂SO₄ 6 g Antifoam 204 5 ml Feed Medium—1 L

Glucose 500 g (NH₄)₂SO₄ 72 g KH₂PO₄ 13.5 g MgSO₄ 5 g Inositol 70 mg Thiamine 10 mg Trace Metal Solution 40 ml Vitamins Solution 40 ml Fermentation Parameters: pH 5.5, controlled Temp - 30° C. Air Flow - 1.4 lpm Agitation - 1150 rpm Inoculum - 200 ml overnight culture grown in YEP + 5% glucose + 2.5% olive oil

Example 5D 2 Liter Fed-Batch of Canthaxanthin Producing Strain MF838

FIG. 9e depicts the production and intracellular accumulation of canthaxanthin and echinenone by strain MF838 (Example 2g) in fed-batch fermentation together with DCW levels. This example demonstrates the advantage of a two phase feeding protocol in which the first phase of feeding is designed to maintain excess carbon and oxygen limited conditions, while the second phase of feeding results in oxygen excess conditions via carbon limitation.

Fermentations A and B are depicted on FIG. 9e . Medium and process parameters are described below. In both fermentations, feeding of the glucose containing feed medium was initiated during the early exponential growth phase at a rate of 22.8 ml/hr. In fermentation A, this feed rate continued until the dissolved oxygen reached 20%, at which time feed was added to maintain the dO2 at 20% (DO controlled feed) for the remainder of the fermentation. In fermentation B, the constant feed rate was maintained such that glucose was in excess, and dO2 level was essential zero, until approximately hour 40 of the fermentation. At that time feed was added to maintain the dO2 at 20% (DO controlled feed) for the remainder of the fermentation. As seen in FIG. 9e , the extended period of carbon excess and oxygen limitation resulted in higher peak DCW, altered kinetics of canthaxanthin production, and produced a higher final level canthaxanthin—over 3.5% of DCW.

Batch Medium—1 L

Glucose 40 g Yeast Nitrogen Base 8.2 g w/o Amino Acids and (NH₄)₂SO₄ (NH₄)₂SO₄ 6 g Antifoam 204 5 ml Feed Medium—1 L

Glucose 500 g (NH₄)₂SO₄ 72 g KH₂PO₄ 13.5 g MgSO₄ 5 g Inositol 70 mg Thiamine 10 mg Trace Metal Solution 40 ml Vitamins Solution 40 ml Fermentation Parameters: pH 5.5, controlled Temp - 30° C. Air Flow - 1.4 lpm Agitation - 1150 rpm Inoculum - 200 ml overnight culture grown in YEP + 5% glucose

Example 6 Introduction of Heterologous Carotene Hydroxylase and Carotene Ketolase Genes into Y. lipolytica Strains Producing Carotenoid for Production of Astaxanthin

For introduction of carotene hydroxylase and carotene ketolase into carotenoid producing Y. lipolytica, pMB4692 and pMB4698, described as in Example 1E and 1F above, can be sequentially introduced into MF740 or MF746 (described in Example 2E). For the introduction of pMB4692, the plasmid may be cleaved with SalI or BsrGI to direct integration at the ura3 locus, or with XbaI to promote random integration, selecting for uracil prototrophy. Ura⁺ transformants from MF740 or MF746 harboring pMB4692 are screened for zeaxanthin production in YPD. Zeaxanthin-producing cells are transformed with pMB4698 (which can be cleaved with PpuMI, SspI or BbvCI to direct integration at the ade1 locus, or with EcoRV to promote random integration) and prototrophic colonies are screened for astaxanthin production.

Alternatively, the order of plasmid transformation may be reversed wherein pMB4698 is transformed first and transformants are selected for adenine prototrophy. Ade⁺ transformants from MF740 or MF746 harboring pMB4698 are screened for canthaxanthin production. Canthaxanthin-producing MF740[pMB4698] or MF746[pMB4698] cells are transformed with pMB4692 and prototrophic colonies are screened for astaxanthin production.

In another approach, the carotenoid ketolase and carotenoid hydroxylase genes from P. marcusii can be introduced into a Leu2⁻ version of MF740 or MF746, in order to convert β-carotene into astaxanthin. P. marcusii genomic DNA is amplified with two primers.

CrtZfwd: (SEQ ID NO: 87) 5′ CACACCGTCTCAAatgaccaatttcctgatcgtcgtc CrtZrev: (SEQ ID NO: 88) 5′ CACACAGATCtcacgtgcgctcctgcgcc, and the resulting fragment is cleaved with BsmBI, modified with the Klenow fragment of DNA polymerase, and cleaved with BglII. This fragment is inserted into PmlI- and BamHI-cleaved pINA1269 (J. Mol. Microbiol. Biotechnol. 2 (2000): 207-216), containing the hp4d promoter, the XPR2 terminator, the selectable LEU2 gene, and sequences necessary for selection and propagation in E. coli. The resulting plasmid “pA” contains sequences encoding carotene hydroxylase from P. marcusii (crtZ gene) (Genbank accession: CAB56060.1) under the control of the hp4d promoter.

“pYEG1TEF” is modified by substituting the LIP2 terminator for the XPR2 terminator as follows. pINA1291 is digested with AvrII, modified with the Klenow fragment of DNA polymerase, and cleaved with EcoRI, and the small LIP2t containing fragment is ligated to “pYEG1TEF” that has been digested with SacII, modified with T4 DNA polymerase in the presence of dNTP, and cleaved with EcoRI. The resulting plasmid is named “pYEG1TEF-LIP2t”.

In order to amplify the carotenoid ketolase gene, P. marcusii genomic DNA is amplified with two primers.

CrtWfwd: (SEQ ID NO: 89) 5′ CACACCCTAGGCCatgagcgcacatgccctgc CrtWrev: (SEQ ID NO: 90) 5′ CACACAAGCTTtcatgcggtgtcccccttg, and the resulting fragment is cleaved with AvrII and HindIII, and inserted into AvrII- and HindIII-cleaved “pYEG1TEF-LIP2t”. The resulting plasmid, “pBt”, contains sequences encoding the carotene ketolase (crtW gene) (Genbank accession: CAB56059.1) under the control of the constitutive TEF1 promoter.

In order to combine the two expression cassettes into a single plasmid, “pBt” is cleaved with ClaI, modified with the Klenow fragment of DNA polymerase, and cleaved with EcoRI, and the crtW-containing fragment is isolated, mixed with the phosphorylated oligonucleotide adaptor pair:

(SEQ ID NO: 91) 5′ AATTCGCGGCCGCT and (SEQ ID NO: 92) 5′ AGCGGCCGCG, cleaved with Nod, and ligated to NotI-digested “pA”. The resulting plasmid, “pABt”, contains both the TEF1p/crtW/LIP2t cassette and the hp4d/crtZ/XPR2t cassette as well as the selectable LEU2 gene.

“pABt” can be introduced into MF740 or MF746 and transformants selected for leucine prototrophy.

Example 7 Partial Inactivation of Y. Lipolytica ERG9 Gene Encoding Squalene Synthase Results in Increased Carotenoid Production

7A.

In order to partially inactivate the ERG9 gene encoding squalene synthase, the neighboring FOL3 gene is disrupted, resulting in a folinic acid requirement. This strain is then transformed with a mutagenized fragment of DNA partially spanning the two genes, and Fol⁺ transformants are screened for decreased squalene synthase activity.

The following oligonucleotides are synthesized:

PRIMER K (SEQ ID NO: 93) 5′-CCTTCTAGTCGTACGTAGTCAGC; PRIMER L (SEQ ID NO: 94) 5′-CCACTGATCTAGAATCTCTTTCTGG and used to amplify a 2.3 kb fragment from Y. lipolytica genomic DNA spanning most of the FOL3 gene, using Pfu polymerase. The resulting fragment is cleaved with XbaI and phosphorylated, then ligated into pBluescriptSK⁻ that has been cleaved with KpnI, treated with T4 DNA polymerase (T4pol) in the presence of dNTPs, and subsequently cleaved with XbaI. The resultant plasmid, designated pBS-fol3, is then cleaved with Acc65I and EcoRI, treated with T4pol as above, and ligated to the 3.4 kb EcoRV-SpeI ADE1 fragment (treated with T4pol) from pMB4529.

The resulting plasmid, pBSfol3Δade, can be cleaved with BsiWI and XbaI to liberate a 5.5 kb fragment that is used to transform MF740 or MF746 to adenine prototrophy. Resulting Ade⁺ transformants are screened for a folinic acid requirement, and for homologous integration by PCR analysis.

Strains that harbor the resultant fol3ΔADE1 allele can be transformed with a 3.5 kb DNA fragment generated by mutagenic PCR amplification using the primers:

PRIMER M (SEQ ID NO: 95) 5′-GGCTCATTGCGCATGCTAACATCG; PRIMER N (SEQ ID NO: 96) 5′-CGACGATGCTATGAGCTTCTAGACG, and Y. lipolytica genomic DNA as template. The resulting fragment containing the N-terminal three-quarters of the FOL3 ORF and the C-terminal nine-tenths of the ERG9 ORF is used to transform strains. The resulting Fol⁺ Ade⁻ transformants are screened for decreased squalene synthase activity by sensitivity to agents such as zaragozic acid, itraconazole, or fluconazole. Additionally, the resulting transformants are screened for increased carotenoid production.

7B.

Alternatively, the PCR fragment produced in 7A could be cloned and altered in such a way as to remove the 3′-untranslated region of ERG9 gene. Replacement of the fol3ΔADE1 disruption by this fragment results in decreased expression of squalene synthase [Schuldiner et al. (2005), Cell 123:507-519][Muhlrad and Parker (1999), RNA 5:1299-1307], which can be confirmed as in 7A. This approach may also be used in a Fol⁺ Ade⁻ strain, using the ADE1 marker to disrupt the ERG9 3′-UTR.

7C.

In still another approach, partially defective ERG9 alleles can be identified in S. cerevisiae using plasmid shuffling techniques [Boeke et al. (1987), Methods Enzymol. 154:164-175], and using drug sensitivities as a phenotype. Defective genes can be transferred to Y. lipolytica using standard molecular genetic techniques.

Example 8 Treatment of Y. Lipolytica Strains Producing Carotenoid with Inhibitor of an Isoprenoid Biosynthesis Competitor Polypeptide Results in Increased Carotenoid Production

Cultures produced in Example 2 are treated with the squalene synthase inhibitor, zaragozic acid (zaragozic acid at 0.5 μM) and monitored for β-carotene production, as described above.

Example 9 Constructing an Oleaginous Strain of Saccharomyces cerevisiae

The genes encoding the two subunits of ATP-citrate lyase from N. crassa, the AMP deaminase from Saccharomyces cerevisiae, and the cytosolic malic enzyme from M. circinelloides are overexpressed in S. cereviseae strains in order to increase the total lipid content. Similar approaches to enhance lipid production could be employed in other host organisms such as Xanthophyllomyces dendrorhous (Phaffia rhodozyma), using the same, homologous, or functionally similar oleaginic polypeptides.

Qiagen RNAEasy kits (Qiagen, Valencia, Calif.) are used to prepare messenger RNA from lyophilized biomass prepared from cultures of N. crassa. Subsequently, RT-PCR is performed in two reactions containing the mRNA template and either of the following primer pairs.

acl1: 1fwd: (SEQ ID NO: 97) 5′ CACACGGATCCTATAatgccttccgcaacgaccg 1rev: (SEQ ID NO: 98) 5′ CACACACTAGttaaatttggacctcaacacgaccc acl2: 2fwd: (SEQ ID NO: 99) 5′ CACACGGATCCAATATAAatgtctgcgaagagcatcctcg 2rev: (SEQ ID NO: 100) 5′ CACACGCATGCttaagcttggaactccaccgcac

The resulting fragment from the acl1 reaction is cleaved with SpeI and BamHI, and that from the acl2 reaction is cleaved with BamHI and SphI, and both are ligated together into YEp24 that has been digested with NheI and SphI, creating the plasmid “p12”. The bi-directional GAL1-10 promoter is amplified from S. cerevisiae genomic DNA using the primers.

gal10: (SEQ ID NO: 101) 5′ CACACGGATCCaattttcaaaaattcttactttttttttggatggac gal1: (SEQ ID NO: 102) 5′ CACACGGATCCttttttctccttgacgttaaagtatagagg, and the resulting 0.67 kb fragment is cleaved with BamHI and ligated in either orientation to BamHI-digested “p12” to create “p1gal2” and “p2gal1”, containing GAL1-acl1/GAL10-acl2 and GAL10-acl1/GAL1-acl2, respectively (Genbank accession: acl1: CAB91740.2; acl2: CAB91741.2).

In order to amplify the S. cereviseae gene encoding AMP deaminase and a promoter suitable for expressing this gene, S. cerevisiae genomic DNA is amplified using two primer pairs in separate reactions:

AMD1 ORF: AMD1FWD: (SEQ ID NO: 103) 5′ CACACGAGCTCAAAAatggacaatcaggctacacagag AMD1rev: (SEQ ID NO: 104) 5′ CACACCCTAGGtcacttttcttcaatggttctcttgaaattg GAL7p: gal7prox: (SEQ ID NO: 105) 5′ CACACGAGCTCggaatattcaactgtttttttttatcatgttgatg gal7dist: (SEQ ID NO: 106) 5′ CACACGGAtccttcttgaaaatatgcactctatatcttttag, and the resulting fragment from the AMD1 reaction (2.4 kb) is cleaved with SacI and AvrII, and that from the GAL7 reaction (0.7 kb) is cleaved with BamHI and SphI, and both are ligated together into YEp13 that has been digested with NheI and BamHI, creating the plasmid “pAMPD”. This plasmid carries the S. cerevisiae gene, AMD1, encoding AMP deaminase, under the control of the galactose-inducible GAL7 promoter.

Messenger RNA is prepared from lyophilized biomass of M. circinelloides, as described above, and the mRNA template is used in a RT-PCR reaction with two primers:

MAEfwd: (SEQ ID NO: 107) 5′ CACACGCTAGCTACAAAatgttgtcactcaaacgcatagcaac MAErev: (SEQ ID NO: 108) 5′ CACACGTCGACttaatgatctcggtatacgagaggaac, and the resulting fragment is cleaved with NheI and SalI, and ligated to XbaI- and XhoI-digested pRS413TEF (Mumberg, D. et al. (1995) Gene, 156:119-122), creating the plasmid “pTEFMAE”, which contains sequences encoding the cytosolic NADP⁺-dependant malic enzyme from M. circinelloides (E.C. 1.1.1.40; mce gene; Genbank accession: AY209191) under the control of the constitutive TEF1 promoter.

The plasmids “p1gal2”, “pAMPD”, and “pTEFMAE” are sequentially transformed into a strain of S. cereviseae to restore prototrophy for uracil (“p1gal2”), leucine (“pAMPD”), and histidine (“pTEFMAE”) (Guthrie and Fink Methods in Enzymology 194:1-933, 1991). The resulting transformants are tested for total lipid content following shake flask testing in either synthetic complete (SC) medium lacking uracil, leucine and histidine, as described in Example 3, or in a 2-step fermentation process. In the 2-step process, 1.5 ml of cells from an overnight 2 ml roll tube culture containing SC medium lacking uracil, leucine and histidine are centrifuged, washed in distilled water, and resuspended in 20 ml of a nitrogen-limiting medium suitable for lipid accumulation (30 g/L glucose, 1.5 g/L yeast extract, 0.5 g/L NH₄C1, 7 g/L KH₂PO₄, 5 g/L Na₂HPO₄-12H₂O, 1.5 g/L MgSO₄-7H₂O, 0.08 g/L FeCl₃-6H₂O, 0.01 g/L ZnSO₄-7H₂O, 0.1 g/L CaCl₂-2H₂O, 0.1 mg/L MnSO₄-5H₂O, 0.1 mg/L CuSO₄-5H₂O, 0.1 mg/L Co(NO₃)₂-6H₂O; pH 5.5 (J Am Oil Chem Soc 70:891-894 (1993)).

Intracellular lipid content of the modified and control S. cerevisiae strains is analyzed using the fluorescent probe, Nile Red (J Microbiol Meth (2004) 56:331-338). In brief, cells diluted in buffer are stained with Nile Red, excited at 488 nm, and the fluorescent emission spectra in the wavelength region of 400-700 nm are acquired and compared to the corresponding spectra from cells not stained with Nile Red. To confirm results from the rapid estimation method, the total lipid content is determined by gas chromatographic analysis of the total fatty acids directly transmethylesterified from dried cells, as described (Appl Microbiol Biotechnol. 2002 November; 60(3):275-80). Non-transformed S. cerevisiae strains produce 6% and 10% total lipid (dry cell weight basis) after growth in YPD and lipid accumulation medium, respectively. Yeast strains expressing the multiple oleaginic polypeptides produce 17% and 25% total lipid following growth in YPD and lipid accumulation medium, respectively.

Example 10 Introduction of Heterologous Carotene Hydroxylase into Y. Lipolytica Strains Producing Carotenoid or Production of Zeaxanthin

MF578 (tef-carRP tef-carB) was transformed with pMB4692 that had been cleaved with SalI. Several Ura⁺ colonies inferred to contain tef-crtZ by PCR analysis were able to produce zeaxanthin in YPD shake flasks, and in one case, all of the β-carotene was depleted.

Example 11 Regulatory Sequences

Sequences which consist of, consist essentially of, and comprise the following regulatory sequences (e.g. promoters and terminator sequences, including functional fragments thereof) may be useful to control expression of endogenous and heterologous genes in engineered host cells, and particularly in engineered fungal cells described herein.

Met2 promoter (SEQ ID NO: 109) 5′cctctcactttgtgaatcgtgaaacatgaatcttcaagccaagaatgttaggcaggggaagctttctttcagactttttggaattggtcctcttttggac attattgacgatattattattttttccccgtccaatgttgacccttgtaagccattccggttctggagcgcatctcgtctgaaggagtcttcgtgtggctata actacaagcgttgtatggtggatcctatgaccgtctatatagggcaacttttgctcttgttcttccccctccttgagggacgtatggcaatggctatgaca actatcgtagtgagcctctataacccattgaagtacaagtcctccaccttgctgccaaactcgcgagaaaaaaagtccaccaactccgccgggaaat actggagaacacctctaagacgtgggcttctgcacctgtgtggcttgggtctgggttttgcgagctctgagccacaacctaaggacggtgtgattgg gagataagtagtcgttggttttctaatcgcacgtgatatgcaagccacacttataacacaatgaagacaggccgatgaactgcatgtcattgtacaggt gcggagagcaagaaactctggggcggaggtgaaagatgagacaaaaagcctcaggtgcaaggtagggagttgatcaacgtcaaacacaaataa tctaggttgttaggcagctaaacatgtatataactgggctgccaccgagtgttacttgtcattaacgtcgcattttcgcctacacaaaatttgggttactcg ccactacactgctcaaatctttcagctgtgcaacaagctttcaggtcacacatagactcgcataaggacccgggtcatctgttattctccactggtaaac caatagtcctagctgatttgggtacagaagctcactttcacatcttttcatcttcttctacaaccatc Met3 promoter (SEQ ID NO: 110) 5′atctgtgaggagcccctggcgtcactgtcgactgtgccggcatttctgatggtatttccagccccgcagttctcgagacccccgaacaaatgtgcc acacccttgccaaaatgacgaatacacggcgtcgcggccgggaatcgaactcttggcaccgccacaggagtgaaatttgaaatttgaaatttgaaa aataattcacattttgagtttcaataatatatcgatgaccctcccaaaagacccaagtcgagacgcaaaaaaacacccagacgacatggatgcggtc acgtgaccgcaaaaaccgccccggaaatccgtttgtgacgtgttcaattccatctctatgtttttctgcggtttctacgatgccgcaatggtggccaatg tgcgtttcactgccgtagtggctggaacaagccacagggggtcgtcgggccaatcagacggtccctgacatggttctgcgccctaacccgggaac tctaacccccgtggtggcgcaatcgctgtcttcatgtgctttatctcacgtgacggctggaatctggcagaagacggagtatgtacattttgtcgttggt cacgttatccctaaaacgtggtgtttaaactggtcgaatgcttggcccagaacacaagaagaaaaaaacgagacaacttgatcagtttcaacgccac agcaagcttgtcttcactgtggttggtcttctccacgccacaagcaacacgtacatgtcaattacgtcagggtcttttaagttctgtggcttttgaaccagt tataaagaaccaaccacccttttttcaaagctaatcaagacggggaaattttttttttgatatttttcgaca Met6 promoter (SEQ ID NO: 111) 5′gatactgcagacggtgcattacttacccgtgtcgactgagagtctacttggtacttggccctgtggctaagcagtatttgagcaacaatgcaatgca gttgctgactcggttccagatccccttgccccgatgtgtggaagcgttgtttttggggcaagggcatgtgggggctgcatcatactgtggctggggcc gttggaagagccgtcggcagcgagcctgagtcgcttctcggggccttattccccccgcctctaggtcagcggcggccgaagtgtcgtactcagctc gcctgtacagtatgacgtgaccgaatagcctctggaaggttggagaagtacagtgcaaaaaaaagttgcaaaatttcattttagcgttcgatccgacg tggcagttggacaatgaatcgatggagacatgatcatgggcagaaatagaaggtctccatgttcaatggcagtaccaattgagcaacagacgggtc gacaggcggcgggcacaccatccgccctccacatggcgcaatcgtcagtgcagcgattcgtactcggattgcatcatgttgcaccgaaagttggg gcccgcacgttggagaggcgaggagccagggttagctttggtggggtcctttgttgtcacgtggcatcagcgaatggcgtcctccaatcagggcc gtcagcgaagtcggcgtgtgatagtgcgtggggagcgaatagagtttctgggggggggcggcccaaaacgtgaaatccgagtacgcatgtaga gtgtaaattgggtgtatagtgacattgtttgactctgaccctgagagtaatatataatgtgtacgtgtccccctccgttggtcttctttttttct cctttctcctaaccaacacccaaactaatcaatc Met25 promoter (SEQ ID NO: 112) 5′aagtcgtattaacataactttccttacatttttttaaagcacgtcactatccacgtgacctagccacgcgataccaagtattcatccataatgacacact catgacgtccggaggacgtcatcatcgtccagtcacgtgccaaggcacatgactaatcataacaccttatgactagcttctgaatcgctacacagttc caattcgcaaataaactcgaaatgacgaaatgccataataaaaatgacgaaactcgagattgagagcagcacatgcactgaagtggtggacaacc agcgtatccggagacacgacggatccagcaccatggaagctggccgaaaaagagatccccagcacattgagcaaccaagtcagctcaattgagt aacatcacacactcagatcgagtctgatggtggtccccttttgttccttcacttgaaaaataattgaaaataacaataacaataaaaataaaaacaaaat aaaaataaaaataaaaataaaaataaaaaaataaaaaaaccttgccgcatttagcgtcagccaccccccgcattgacctgagtacgttggattgacc ccgatcctgcacgtcgagcgtggtcggccaaaaagcgcccgtggctggtgagtcagaaatagcagggttgcaagagagagctgcgcaacgagc aataaacggtgtttttttcgcttctgtgctgcttagagtggagagccgaccctcgccatgctcacgtgaccattcacgtggttgcaaactccaccttagt atagccgtgtccctctcgctacccattatcgcatcgtactccagccacatttttttgttccccgctaaatccggaaccttatctgggtcacgtgaaattgc aatctcgacaggaggttatacttatagagtgagacactccacgcaaggtgttgcaagtcaattgacaccacctcacctcagactaacatccaca Pox2 promoter (SEQ ID NO: 113) 5′gaatctgcccccacattttatctccgcttttgactgtttttctcccccctttcacactctgcttttggctacataaaccccgcaccgtttggaactct gttggtccggggaagccgccgttaggtgtgtcagatggagagcgccagacgagcagaaccgagggacagcggatcgggggagggctgtcacgtga cgaagggcactgttgacgtggtgaatgtcgcccgttctcacgtgacccgtctcctctatatgtgtatccgcctctttgtttggttttttttctgcttcccccc ccccccccccaccccaatcacatgctcagaaagtagacatctgcatcgtcctgcatgccatcccacaagacgaacaagtgataggccgagagccg aggacgaggtggagtgcacaaggggtaggcgaatggtacgattccgccaagtgagactggcgatcgggagaagggttggtggtcatgggggat agaatttgtacaagtggaaaaaccactacgagtagcggatttgataccacaagtagcagagatatacagcaatggtgggagtgcaagtatcggaat gtactgtacctcctgtactcgtactcgtacggcactcgtagaaacggggcaatacgggggagaagcgatcgcccgtctgttcaatcgccacaagtc cgagtaatgctcgagtatcgaagtcttgtacctccctgtcaatcatggcaccactggtcttgacttgtctattcatactggacaagcgccagagttaagc ttgtagcgaatttcgccctcggacatcaccccatacgacggacacacatgcccgacaaacagcctctcttattgtagctgaaagtatattgaatgtgaa cgtgtacaatatcaggtaccagcgggaggttacggccaaggtgataccggaataaccctggcttggagatggtcggtccattgtactgaagtgtccgtgtc gtttccgtcactgccccaattggacatgtttgtttttccgatctttcgggcgccctctccttgtctccttgtctgtctcctggactgttgctaccccatt tctttggcctccattggttcctccccgtctttcacgtcgtctatggttgcatggtttcccttatacttttccccacagtcacatgttatggaggggtctag atggaggcctaattttgacgtgcaaggggcgaattggggcgagaaacacgtcgtggacatggtgcaaggcccgcagggttgattcgacgcttttccgc gaaaaaaacaagtccaaatacccccgtttattctccctcggctctcggtatttcacatgaaaactataacctagactacacgggcaaccttaaccccag agtatacttatataccaaagggatgggtcctcaaaaatcacacaagcaacg Yef3 (YALI0E13277g) promoter (SEQ ID NO: 114) 5′cgccattcggttccttccagaccattccagatcaatccacctcttcttatctcaggtgggtgtgctgacatcagaccccgtagcccttctcccagtgg cgaacagcaggcataaaacagggccattgagcagagcaaacaaggtcggtgaaatcgtcgaaaaagtcggaaaacggttgcaagaaattggag cgtcacctgccaccctccaggctctatataaagcattgccccaattgctaacgcttcatatttacacctttggcaccccagtccatccctccaataaaat gtactacatgggacacaacaagagaggatgcgcgcccaaaccctaacctagcacatgcacgatgattctctttgtctgtgaaaaaatttttccaccaa aatttccccattgggatgaaaccctaaccgcaaccaaaagtttttaactatcatcttgtacgtcacggtttccgattcttctcttctctttcatcatcat cacttgtgacc Cam1 (YALI0C24420g) promoter (SEQ ID NO: 115) 5′aactaccataaagtaccgagaaatataggcaattgtacaaattgtccacctccttcacttacattaccgaaccatggccatatcaccaaaatacccc gagtgctaaaacacctccctccaaatgttctcttaccttccaccgaaaaccgatcttattatcccaacgcttgttgtggcttgacgcgccgcacccgctg ggcttgccatttcgataccaatccaagaggaaaagctcatgagaaacaatcggaatatcacgagaacggcctggcgaaccaacaggatatttttga atataattacccctcgaatctagtcatatctatgtctactgtagacttgggcggcatcatgatgtacattattttagcgtctggaaccctaaagttcacgta caatcatgtgacaaacgaggctaaaaaatgtcaatttcgtatattagtgttattacgtggctcacatttccgaatcatctaccaccccccacctaaaaa YALI0D16467g promoter (SEQ ID NO: 116) 5′tttttttaattttcatatttattttcatatttattttcatatttattttcatttatttattcatgtatttatttattactttttaagtattttaaa ctcctcactaaaccgtcgattgcacaatattaaccttcattacacctgcagcgtggtttttgtggtcgttagccgaagtcttccaacgtg ggtataagtaggaacaattgggccgattttttgagccgtctaaatctctcgactcaattgatctgctgtcgaaaatccggctctctagc tccttttccccgtccgctggagctcctcttcattgtgccgtttttccaacatttaactttgccacccaccaccacccccactaccatcacc cactcgatctctgttcgtgtcaccacgactttgtcttctcacacatactctgtttgtgcaccacacattgcgaa Tef4 (YALI0B12562g) promoter (SEQ ID NO: 117) 5′gctacaatagctttattggccctattgagcacgctacaattcggtccagtatgtacaacgtctatgcgcactaacggccatacagtgagttacagca cacccaaaagtaaccctgcctgacctgtctgcctgagacaggaagattaactcttgtagtgaccgagctcgataagactcaagccacacaattttttta tagccttgcttcaagagtcgccaaaatgacattacacaactccacggaccgtcggttccatgtccacacccttggatgggtaagcgctccacgcacg taccacgtgcattgagtttaaccacaaacataggtctgtgtcccagagttaccctgctgcatcagccaagtcttgaaagcaaaatttcttgcacaattttt cctcttcttttcttcactgatcgcagtccaaacacaaaca YALI0D12903g promoter (SEQ ID NO: 118) 5′gcgctctgatccacttgtatggctccaagttcagtgtaccaagtagttggtgatgcagggagggatgtctctatccaccaataatgaactcatgggcgaa attgtttctgttaaacactccaactgtcgttttaaatctcattctctttgcatttggactccattcgcttccgttgggccaatataatccatcgtaacgtac tttagatggaaatttagttacctgctacttgtctcaacaccccaacaggggctgttcgacagaggtaatagagcgtcaatgggttaataaaaacacactgtcg attttcactcattgtctttatgatattacctgttttccgctgttatcaatgccgagcatcgtgttatatcttccaccccaactacttgcatttacttaactat tacctcaactatttacaccccgaattgttacctcccaataagtaactttatttcaaccaatgggacgagagcatctctgagaacatcgatctatctctgtca atattgcccagaatcgttcgaaaaaaaacaccaaaaggtttacagcgccattataaatataaattcgttgtcaattcccccgcaatgtctgttgaaatctc attttgagaccttccaacattaccctctctcccgtctggtcacatgacgtgactgcttcttcccaaaacgaacactcccaactcttcccccccgtcagtg aaaagtatacatccgacctccaaatcttttcttcactcaac Tef1 (YALI0C09141g) promoter (SEQ ID NO: 119) 5′agagacgggttggcggcgtatttgtgtcccaaaaaacagccccaattgccccaattgaccccaaattgacccagtagcgggcccaaccccggc gagagcccccttcaccccacatatcaaacctcccccggttcccacacttgccgttaagggcgtagggtactgcagtctggaatctacgcttgttcaga ctttgtactagtttctttgtctggccatccgggtaacccatgccggacgcaaaatagactactgaaaatttttttgctttgtggttgggactttagccaagg gtataaaagaccaccgtccccgaattacctttcctcttcttttctctctctccttgtcaactcacacccgaaatcgttaagcatttccttctgagtataa gaatcattc Fba1 (YALI0E26004g) promoter (SEQ ID NO: 120) 5′gctgcgctgatctggacaccacagaggttccgagcactttaggttgcaccaaatgtcccaccaggtgcaggcagaaaacgctggaacagcgtg tacagtttgtcttagcaaaaagtgaaggcgctgaggtcgagcagggtggtgtgacttgttatagcctttagagctgcgaaagcgcgtatggatttggc tcatcaggccagattgagggtctgtggacacatgtcatgttagtgtacttcaatcgccccctggatatagccccgacaataggccgtggcctcatttttt tgccttccgcacatttccattgctcggtacccacaccttgcttctcctgcacttgccaaccttaatactggtttacattgaccaacatcttacaagcgggg ggcttgtctagggtatatataaacagtggctctcccaatcggttgccagtctcttttttcctttctttccccacagattcgaaatctaaactacacatc Pox2 terminator: (SEQ ID NO: 121) 5′gatgaggaatagacaagcgggtatttattgtatgaataaagattatgtattgattgcaaaaaagtgcatttgtagatgtggtttattgtagagagtacg gtatgtactgtacgaacattaggagctacttctacaagtagattttcttaacaagggtgaaatttactaggaagtacatgcatatttcgttagtagaatcac aaaagaaatgtacaagcacgtactacttgtactccacaatgtggagtgggagcaaaaaaattggacgacaccggaatcgaaccggggacctcgc gcatgctaagcgcatgtgataaccaactacaccagacgcccaagaactttcttggtgattatggaatacgtggtctgctatatctcaattttgctgtaatg aatcattagaattaaaaaaaaaaccccatttttgtgtgattgtcggccaagagatggaacaggaagaatacgtgaacaagcgagcacgaatgccatatg ctcttctgaacaaccgagtccgaatccgatttgtgggtatcacatgtctcaagtagctgaaatgtatttcgctagaataaaataaatgagattaagaattaa aaatattggaatatattttcctagaatagaaactttggattttttttcggctattacagtctgaactggacaaacggctgactatatataaatattattggg tctgttttcttgtttatgtcgaaattatctgggttttactactgtgtcgtcgagtatagagtggcctgactggagaaaatgcagtagtatggacagtaggt actgccagccagagaagtttttggaattgatacttgagtcatttttccattccccattccccattccaacacaatcaactgtttctgaacattttccaaaacg cggagatgtatgtcacttggcactgcaagtctcgattcaaaatgcatctctttcagaccaaagtgtcatcagctttgtttggccccaaattaccgcaaat acttgtcgaaattgaagtgcaatacggcctcgtctgccatgaaacctgcctattctcttcaaattggcgtcaggtttcacgtccagcattcctcgcccag acagagttgctatggttgaatcgtgtactgttaatatatgtatgtattatactcgtactacgatatactgttcaatagagtctcttataatcgtacga cgattctgggca

Example 12 Y. lipolytica Oleaginic and Isoprenoid Biosynthesis Genes

FIG. 10 is a list of Y. lipolytica genes representing various polypeptides (e.g. oleaginic and isoprenoid biosynthesis peptides) useful in the fungal strains and methods described herein. The Genbank accession number and GI number is given for each polypeptide in addition to oligo pairs which can be used to amplify the coding region for each gene from Y. lipolytica genomic DNA or cDNA. Resulting PCR fragments can be cleaved with restriction enzyme pairs (e.g. depending on what site is present within the oligo sequence, XbaI/MluI or NheI/MluI or XbaI/AscI or NheI/AscI) and inserted into expression vectors (e.g. fungal expression vectors including Y. lipolytica expression vectors such as MB4629 and MB4691 described herein).

The DNA and proteins they encode of the Y. lipolytica genes represented in FIG. 10 are as follows (intron sequence is underlined):

YALI0F30481g DNA: (SEQ ID NO: 122) atgtcgcaaccccagaacgttggaatcaaagccctcgagatctacgtgccttctcgaattgtcaaccaggctgagctcgagaagcacgacggtgtc gctgctggcaagtacaccattggtcttggtcagaccaacatggcctttgtcgacgacagagaggacatctattcctttgccctgaccgccgtctctcg actgctcaagaacaacaacatcgaccctgcatctattggtcgaatcgaggttggtactgaaacccttctggacaagtccaagtccgtcaagtctgtgc tcatgcagctctttggcgagaacagcaacattgagggtgtggacaacgtcaacgcctgctacggaggaaccaacgccctgttcaacgctatcaact gggttgagggtcgatcttgggacggccgaaacgccatcgtcgttgccggtgacattgccctctacgcaaagggcgctgcccgacccaccggagg tgccggctgtgttgccatgctcattggccccgacgctcccctggttcttgacaacgtccacggatcttacttcgagcatgcctacgatttctacaagcct gatctgacctccgagtacccctatgttgatggccactactccctgacctgttacacaaaggccctcgacaaggcctacgctgcctacaacgcccgag ccgagaaggtcggtctgttcaaggactccgacaagaagggtgctgaccgatttgactactctgccttccacgtgcccacctgcaagcttgtcaccaa gtcttacgctcgacttctctacaacgactacctcaacgacaagagcctgtacgagggccaggtccccgaggaggttgctgccgtctcctacgatgc ctctctcaccgacaagaccgtcgagaagaccttccttggtattgccaaggctcagtccgccgagcgaatggctccttctctccagggacccaccaa caccggtaacatgtacaccgcctctgtgtacgcttctctcatctctctgctgacttttgtccccgctgagcagctgcagggcaagcgaatctctctcttct cttacggatctggtcttgcttccactcttttctctctgaccgtcaagggagacatttctcccatcgtcaaggcctgcgacttcaaggctaagctcgatgac cgatccaccgagactcccgtcgactacgaggctgccaccgatctccgagagaaggcccacctcaagaagaactttgagccccagggagacatca agcacatcaagtctggcgtctactacctcaccaacatcgatgacatgttccgacgaaagtacgagatcaagcagtag Protein: (SEQ ID NO: 123) Msqpqnvgikaleiyvpsrivnqaelekhdgvaagkytiglgqtnmafvddrediysfaltavsrllknnnidpasigrievgtetlldksksvk svlmqlfgensniegvdnvnacyggtnalfnainwvegrswdgrnaivvagdialyakgaarptggagcvamligpdaplvldnvhgsyfe haydfykpdltseypyvdghysltcytkaldkayaaynaraekvglfkdsdkkgadrfdysafhvptcklvtksyarllyndylndkslyegq vpeevaavsydasltdktvektflgiakaqsaermapslqgptntgnmytasvyaslislltfvpaeqlqgkrislfsygsglastlfsltvkgdisp ivkacdfkaklddrstetpvdyeaatdlrekahlkknfepqgdikhiksgvyyltniddmfrrkyeikq YALI0B16038g DNA: (SEQ ID NO: 124) atggactacatcatttcggcgccaggcaaagtgattctatttggtgaacatgccgctgtgtttggtaagcctgcgattgcagcagccatcgacttgcga acatacctgcttgtcgaaaccacaacatccgacaccccgacagtcacgttggagtttccagacatccacttgaacttcaaggtccaggtggacaagc tggcatctctcacagcccagaccaaggccgaccatctcaattggtcgactcccaaaactctggataagcacattttcgacagcttgtctagcttggcg cttctggaagaacctgggctcactaaggtccagcaggccgctgttgtgtcgttcttgtacctctacatccacctatgtcccccttctgtgtgcgaagatt catcaaactgggtagttcgatcaacgctgcctatcggcgcgggcctgggctcttccgcatccatttgtgtctgtttggctgcaggtcttctggttctcaa cggccagctgagcattgaccaggcaagagatttcaagtccctgaccgagaagcagctgtctctggtggacgactggtccttcgtcggtgaaatgtg cattcacggcaacccgtcgggcatcgacaatgctgtggctactcagggaggtgctctgttgttccagcgacctaacaaccgagtccctcttgttgac attcccgagatgaagctgctgcttaccaatacgaagcatcctcgatctaccgcagacctggttggtggagtcggagttctcactaaagagtttggctc catcatggatcccatcatgacttcagtaggcgagatttccaaccaggccatggagatcatttctagaggcaagaagatggtggaccagtctaaccttg agattgagcagggtatcttgcctcaacccacctctgaggatgcctgcaacgtgatggaagatggagctactcttcaaaagttgagagatatcggttcg gaaatgcagcatctagtgagaatcaatcacggcctgcttatcgctatgggtgtttcccacccgaagctcgaaatcattcgaactgcctccattgtccac aacctgggtgagaccaagctcactggtgctggaggaggaggttgcgccatcactctagtcacttctaaagacaagactgcgacccagctggagga aaatgtcattgctttcacagaggagatggctacccatggcttcgaggtgcacgagactactattggtgccagaggagttggtatgtgcattgaccatc cctctctcaagactgttgaagccttcaagaaggtggagcgggcggatctcaaaaacatcggtccctggacccattag Protein: (SEQ ID NO: 125) mdyiisapgkvilfgehaavfgkpaiaaaidlrtyllvetttsdtptvtlefpdihlnfkvqvdklasltaqtkadhlnwstpktldkhifdslsslall eepgltkvqqaavvsflylyihlcppsvcedssnwvvrstlpigaglgssasicyclaagllvlngqlsidqardfksltekqlslvddwsfvgem cihgnpsgidnavatqggallfqrpnnrvplvdipemkllltntkhprstadlvggvgvltkefgsimdpimtsvgeisnqameiisrgkkmv dqsnleieqgilpqptsedacnvmedgatlqklrdigsemqhlvrinhglliamgvshpkleiirtasivhnlgetkltgaggggcaitlvtskdk tatqleenviafteemathgfevhettigargvgmcidhpslktveafkkveradlknigpwth YALI0E06193g DNA: (SEQ ID NO: 126) atgaccacctattcggctccgggaaaggccctcctttgcggcggttatttggttattgatccggcgtattcagcatacgtcgtgggcctctcggcgcgt atttacgcgacagtttcggcttccgaggcctccaccacctctgtccatgtcgtctctccgcagtttgacaagggtgaatggacctacaactacacgaa cggccagctgacggccatcggacacaacccatttgctcacgcggccgtcaacaccgttctgcattacgttcctcctcgaaacctccacatcaacatc agcatcaaaagtgacaacgcgtaccactcgcaaattgacagcacgcagagaggccagtttgcataccacaaaaaggcgatccacgaggtgccta aaacgggcctcggtagctccgctgctcttaccaccgttcttgtggcagctttgctcaagtcatacggcattgatcccttgcataacacccacctcgttca caacctgtcccaggttgcacactgctcggcacagaagaagattgggtctggatttgacgtggcttcggccgtttgtggctctctagtctatagacgttt cccggcggagtccgtgaacatggtcattgcagctgaagggacctccgaatacggggctctgttgagaactaccgttaatcaaaagtggaaggtga ctctggaaccatccttcttgccgccgggaatcagcctgcttatgggagacgtccagggaggatctgagactccaggtatggtggccaaggtgatgg catggcgaaaagcaaagccccgagaagccgagatggtgtggagagatctcaacgctgccaacatgctcatggtcaagttgttcaacgacctgcgc aagctctctctcactaacaacgaggcctacgaacaacttttggccgaggctgctcctctcaacgctctaaagatgataatgttgcagaaccctctcgg agaactagcacgatgcattatcactattcgaaagcatctcaagaagatgacacgggagactggtgctgctattgagccggatgagcagtctgcattg ctcaacaagtgcaacacttatagtggagtcattggaggtgttgtgcctggagcaggaggctacgatgctatttctcttctggtgatcagctctacggtg aacaatgtcaagcgagagagccagggagtccaatggatggagctcaaggaggagaacgagggtctgcggctcgagaaggggttcaagtag Protein: (SEQ ID NO: 127) mttysapgkallcggylvidpaysayvvglsariyatvsaseasttsvhvvspqfdkgewtynytngqltaighnpfahaavntvlhyvpprnl hinisiksdnayhsqidstqrgqfayhkkaihevpktglgssaalttvlvaallksygidplhnthlvhnlsqvahcsaqkkigsgfdvasavcgs lvyrrfpaesvnmviaaegtseygallrttvnqkwkvtlepsflppgisllmgdvqggsetpgmvakvmawrkakpreaemvwrdlnaan mlmvklfildlrklsltnneayeqllaeaaplnalkmimlqnplgelarciitirkhlkkmtretgaaiepdeqsallnkcntysgviggvvpgag gydaisllvisstvnnvkresqgvqwmelkeeneglrlekgfk YALI0F05632g DNA: (SEQ ID NO: 128) atgatccaccaggcctccaccaccgctccggtgaacattgcgacactcaagtactggggcaagcgagaccctgctctcaatctgcccactaacaac tccatctccgtgactttgtcgcaggatgatctgcggaccctcaccacagcctcgtgttcccctgatttcacccaggacgagctgtggctcaatggcaa gcaggaggacgtgagcggcaaacgtctggttgcgtgtttccgagagctgcgggctctgcgacacaaaatggaggactccgactcttctctgcctaa gctggccgatcagaagctcaagatcgtgtccgagaacaacttccccaccgccgctggtctcgcctcatcggctgctggctttgccgccctgatccg agccgttgcaaatctctacgagctccaggagacccccgagcagctgtccattgtggctcgacagggctctggatccgcctgtcgatctctctacgga ggctacgtggcatgggaaatgggcaccgagtctgacggaagcgactcgcgagcggtccagatcgccaccgccgaccactggcccgagatgcg agccgccatcctcgttgtctctgccgacaagaaggacacgtcgtccactaccggtatgcaggtgactgtgcacacttctcccctcttcaaggagcga gtcaccactgtggttcccgagcggtttgcccagatgaagaagtcgattctggaccgagacttccccacctttgccgagctcaccatgcgagactcaa accagttccacgccacctgtctggactcgtatcctcccattttctacctcaacgacgtgtcgcgagcctccattcgggtagttgaggccatcaacaag gctgccggagccaccattgccgcctacacctttgatgctggacccaactgtgtcatctactacgaggacaagaacgaggagctggttctgggtgct ctcaaggccattctgggccgtgtggagggatgggagaagcaccagtctgtggacgccaagaagattgatgttgacgagcggtgggagtccgagc tggccaacggaattcagcgggtgatccttaccaaggttggaggagatcccgtgaagaccgctgagtcgcttatcaacgaggatggttctctgaaga acagcaagtag Protein: (SEQ ID NO: 129) mihqasttapvniatlkywgkrdpalnlptnnsisvtlsqddlrtlttascspdftqdelwlngkqedvsgkrlvacfrelralrhkmedsdsslpk ladqklkivsennfptaaglassaagfaaliravanlyelqetpeqlsivarqgsgsacrslyggyvawemgtesdgsdsravqiatadhwpem raailvvsadkkdtssttgmqvtvhtsplfkervttvvperfaqmkksildrdfptfaeltmrdsnqfhatcldsyppifylndvsrasirvveain kaagatiaaytfdagpncviyyedkneelvlgalkailgrvegwekhqsvdakkidvderweselangiqrviltkvggdpvktaeslinedg slknsk YALI0F04015g DNA: (SEQ ID NO: 130) Atgacgacgtcttacagcgacaaaatcaagagtatcagcgtgagctctgtggctcagcagtttcctgaggtggcgccgattgcggacgtgtccaag gctagccggcccagcacggagtcgtcggactcgtcggccaagctatttgatggccacgacgaggagcagatcaagctgatggacgagatctgtg tggtgctggactgggacgacaagccgattggcggcgcgtccaaaaagtgctgtcatctgatggacaacatcaacgacggactggtgcatcgggc cttttccgtgttcatgttcaacgaccgcggtgagctgcttctgcagcagcgggcggcggaaaaaatcacctttgccaacatgtggaccaacacgtgc tgctcgcatcctctggcggtgcccagcgagatgggcgggctggatctggagtcccggatccagggcgccaaaaacgccgcggtccggaagctt gagcacgagctgggaatcgaccccaaggccgttccggcagacaagttccatttcctcacccggatccactacgccgcgccctcctcgggcccctg gggcgagcacgagattgactacattctgtttgtccggggcgaccccgagctcaaggtggtggccaacgaggtccgcgataccgtgtgggtgtcgc agcagggactcaaggacatgatggccgatcccaagctggttttcaccccttggttccggctcatttgtgagcaggcgctgtttccctggtgggacca gttggacaatctgcccgcgggcgatgacgagattcggcggtggatcaagtag Protein: (SEQ ID NO: 131) mttsysdkiksisyssvaqqfpevapiadvskasrpstessdssaklfdghdeeqiklmdeicvvldwddkpiggaskkcchlmdnindglv hrafsvfmfndrgelllqqraaekitfanmwtntccshplavpsemggldlesriqgaknaavrklehelgidpkavpadkfhfltrihyaapss gpwgeheidyilfvrgdpelkvvanevrdtvwvsqqglkdmmadpklvftpwfrliceqalfpwwdqldnlpagddeirrwik YALI0E05753 DNA: (SEQ ID NO: 132) atgtccaaggcgaaattcgaaagcgtgttcccccgaatctccgaggagctggtgcagctgctgcgagacgagggtctgccccaggatgccgtgc agtggttttccgactcacttcagtacaactgtgtgggtggaaagctcaaccgaggcctgtctgtggtcgacacctaccagctactgaccggcaagaa ggagctcgatgacgaggagtactaccgactcgcgctgctcggctggctgattgagctgctgcaggcgtttttcctcgtgtcggacgacattatggat gagtccaagacccgacgaggccagccctgctggtacctcaagcccaaggtcggcatgattgccatcaacgatgctttcatgctagagagtggcat ctacattctgcttaagaagcatttccgacaggagaagtactacattgaccttgtcgagctgttccacgacatttcgttcaagaccgagctgggccagct ggtggatcttctgactgcccccgaggatgaggttgatctcaaccggttctctctggacaagcactcctttattgtgcgatacaagactgcttactactcc ttctacctgcccgttgttctagccatgtacgtggccggcattaccaaccccaaggacctgcagcaggccatggatgtgctgatccctctcggagagt acttccaggtccaggacgactaccttgacaactttggagaccccgagttcattggtaagatcggcaccgacatccaggacaacaagtgctcctggct cgttaacaaagcccttcagaaggccacccccgagcagcgacagatcctcgaggacaactacggcgtcaaggacaagtccaaggagctcgtcatc aagaaactgtatgatgacatgaagattgagcaggactaccttgactacgaggaggaggttgttggcgacatcaagaagaagatcgagcaggttga cgagagccgaggcttcaagaaggaggtgctcaacgctttcctcgccaagatttacaagcgacagaagtag Protein: (SEQ ID NO: 133) mskakfesvfpriseelvqllrdeglpqdavqwfsdslqyncvggklnrglsvvdtyqlltgkkelddeeyyrlallgwliellqafflvsddim desktrrgqpcwylkpkvgmiaindafmlesgiyillkkhfrqekyyidlvelfhdisfktelgqlvdlltapedevdlnrfsldkhsfivrykta yysfylpvvlamyvagitnpkdlqqamdvliplgeyfqvqddyldnfgdpefigkigtdiqdnkcswlvnkalqkatpeqrqilednygvk dkskelvikklyddmkieqdyldyeeevvgdikkkieqvdesrgfkkevlnaflakiykrqk YALI0E18634g DNA: (SEQ ID NO: 134) atgttacgactacgaaccatgcgacccacacagaccagcgtcagggcggcgcttgggcccaccgccgcggcccgaaacatgtcctcctccagc ccctccagcttcgaatactcgtcctacgtcaagggcacgcgggaaatcggccaccgaaaggcgcccacaacccgtctgtcggttgagggcccca tctacgtgggcttcgacggcattcgtcttctcaacctgccgcatctcaacaagggctcgggattccccctcaacgagcgacgggaattcagactcag tggtcttctgccctctgccgaagccaccctggaggaacaggtcgaccgagcataccaacaattcaaaaagtgtggcactcccttagccaaaaacgg gttctgcacctcgctcaagttccaaaacgaggtgctctactacgccctgctgctcaagcacgttaaggaggtcttccccatcatctatacaccgactca gggagaagccattgaacagtactcgcggctgttccggcggcccgaaggctgcttcctcgacatcaccagtccctacgacgtggaggagcgtctg ggagcgtttggagaccatgacgacattgactacattgtcgtgactgactccgagggtattctcggaattggagaccaaggagtgggcggtattggta tttccatcgccaagctggctctcatgactctatgtgctggagtcaacccctcacgagtcattcctgtggttctggatacgggaaccaacaaccaggag ctgctgcacgaccccctgtatctcggccgacgaatgccccgagtgcgaggaaagcagtacgacgacttcatcgacaactttgtgcagtctgcccga aggctgtatcccaaggcggtgatccatttcgaggactttgggctcgctaacgcacacaagatcctcgacaagtatcgaccggagatcccctgcttca acgacgacatccagggcactggagccgtcactttggcctccatcacggccgctctcaaggtgctgggcaaaaatatcacagatactcgaattctcgt gtacggagctggttcggccggcatgggtattgctgaacaggtctatgataacctggttgcccagggtctcgacgacaagactgcgcgacaaaacat ctttctcatggaccgaccgggtctactgaccaccgcacttaccgacgagcagatgagcgacgtgcagaagccgtttgccaaggacaaggccaatt acgagggagtggacaccaagactctggagcacgtggttgctgccgtcaagccccatattctcattggatgttccactcagcccggcgcctttaacga gaaggtcgtcaaggagatgctcaaacacacccctcgacccatcattctccctctttccaaccccacacgtcttcatgaggctgtccctgcagatctgt acaagtggaccgacggcaaggctctggttgccaccggctcgccctttgacccagtcaacggcaaggagacgtctgagaacaataactgctttgtttt ccccggaatcgggctgggagccattctgtctcgatcaaagctcatcaccaacaccatgattgctgctgccatcgagtgcctcgccgaacaggcccc cattctcaagaaccacgacgagggagtacttcccgacgtagctctcatccagatcatttcggcccgggtggccactgccgtggttcttcaggccaag gctgagggcctagccactgtcgaggaagagctcaagcccggcaccaaggaacatgtgcagattcccgacaactttgacgagtgtctcgcctgggt cgagactcagatgtggcggcccgtctaccggcctctcatccatgtgcgggattacgactag Protein: (SEQ ID NO: 135) mlrlrtmrptqtsvraalgptaaarnmsssspssfeyssyvkgtreighrkapttrlsvegpiyvgfdgirllnlphlnkgsgfplnerrefrlsgllp saeatleeqvdrayqqfkkcgtplakngfctslkfqnevlyyalllkhvkevfpiiytptqgeaieqysrlfrrpegcflditspydveerlgafgdh ddidyivvtdsegilgigdqgvggigisiaklalmtlcagvnpsrvipvvldtgtnnqellhdplylgrrmprvrgkqyddfidnfvqsarrlyp kavihfedfglanahkildkyrpeipcfnddiqgtgavtlasitaalkvlgknitdtrilvygagsagmgiaeqvydnlvaqglddktarqniflm drpgllttaltdeqmsdvqkpfakdkanyegvdtktlehvvaavkphiligcstqpgafnekvvkemlkhtprpiilplsnptrlheavpadly kwtdgkalvatgspfdpvngketsennncfvfpgiglgailsrsklitntmiaaaieclaeqapilknhdegvlpdvaliqiisarvatavvlqak aeglatveeelkpgtkehvqipdnfdeclawvetqmwrpvyrplihvrdyd YALI0E11495g DNA: (SEQ ID NO: 136) atgccgcagcaagcaatggatatcaagggcaaggccaagtctgtgcccatgcccgaagaagacgacctggactcgcattttgtgggtcccatctct ccccgacctcacggagcagacgagattgctggctacgtgggctgcgaagacgacgaagacgagcttgaagaactgggaatgctgggccgatct gcgtccacccacttctcttacgcggaagaacgccacctcatcgaggttgatgccaagtacagagctcttcatggccatctgcctcatcagcactctca gagtcccgtgtccagatcttcgtcatttgtgcgggccgaaatgaaccacccccctcccccaccctccagccacacccaccaacagccagaggacg atgacgcatcttccactcgatctcgatcgtcgtctcgagcttctggacgcaagttcaacagaaacagaaccaagtctggatcttcgctgagcaagggt ctccagcagctcaacatgaccggatcgctcgaagaagagccctacgagagcgatgacgatgcccgactatctgcggaagacgacattgtctatga tgctacccagaaagacacctgcaagcccatatctcctactctcaaacgcacccgcaccaaggacgacatgaagaacatgtccatcaacgacgtca aaatcaccaccaccacagaagatcctcttgtggcccaggagctgtccatgatgttcgaaaaggtgcagtactgccgagacctccgagacaagtacc aaaccgtgtcgctacagaaggacggagacaaccccaaggatgacaagacacactggaaaatttaccccgagcctccaccaccctcctggcacga gaccgaaaagcgattccgaggctcgtccaaaaaggagcaccaaaagaaagacccgacaatggatgaattcaaattcgaggactgcgaaatcccc ggacccaacgacatggtcttcaagcgagatcctacctgtgtctatcaggtctatgaggatgaaagctctctcaacgaaaataagccgtttgttgccatc ccctcaatccgagattactacatggatctggaggatctcattgtggcttcgtctgacggacctgccaagtcttttgctttccgacgactgcaatatctag aagccaagtggaacctctactacctgctcaacgagtacacggagacaaccgagtccaagaccaacccccatcgagacttttacaacgtacgaaag gtcgacacccacgttcaccactctgcctgcatgaaccagaagcatctgctgcgattcatcaaatacaagatgaagaactgccctgatgaagttgtcat ccaccgagacggtcgggagctgacactctcccaggtgtttgagtcacttaacttgactgcctacgacctgtctatcgatacccttgatatgcatgctca caaggactcgttccatcgatttgacaagttcaacctcaagtacaaccctgtcggtgagtctcgactgcgagaaatcttcctaaagaccgacaactaca tccagggtcgatacctagctgagatcacaaaggaggtgttccaggatctcgagaactcgaagtaccagatggcggagtaccgtatttccatctacg gtcggtccaaggacgagtgggacaagctggctgcctgggtgctggacaacaaactgttttcgcccaatgttcggtggttgatccaggtgcctcgact gtacgacatttacaagaaggctggtctggttaacacctttgccgacattgtgcagaacgtctttgagcctcttttcgaggtcaccaaggatcccagtac ccatcccaagctgcacgtgttcctgcagcgagttgtgggctttgactctgtcgatgacgagtcgaagctggaccgacgtttccaccgaaagttccca actgcagcatactgggacagcgcacagaaccctccctactcgtactggcagtactatctatacgccaacatggcctccatcaacacctggagacag cgtttgggctataatacttttgagttgcgaccccatgctggagaggctggtgacccagagcatcttctgtgcacttatctggttgctcagggtatcaacc acggtattctgttgcgaaaggtgcccttcattcagtacctttactacctggaccagatccccattgccatgtctcctgtgtccaacaatgcgctgttcctc acgttcgacaagaaccccttctactcatacttcaagcggggtctcaacgtgtccttgtcatcggatgatcctctgcagtttgcttacactaaggaggctc tgattgaggagtactctgtggctgcgctcatttacaagctttccaacgtggatatgtgtgagcttgctcgaaactcggtactgcaatctggctttgagcg aatcatcaaggagcattggatcggcgaaaactacgagatccatggccccgagggcaacaccatccagaagacaaacgtgcccaatgtgcgtctg gccttccgagacgagactttgacccacgagcttgctctggtggacaagtacaccaatcttgaggagtttgagcggctgcatggttaa Protein: (SEQ ID NO: 137) mpqqamdikgkaksvpmpeeddldshfvgpisprphgadeiagyvgceddedeleelgmlgrsasthfsyaeerhlievdakyralhghl phqhsqspvsrsssfvraemnhpppppsshthqqpedddasstrsrsssrasgrkfnrnrtksgsslskglqqlnmtgsleeepyesdddarls aeddivydatqkdtckpisptlkrtrtkddmknmsindvkittttedplvaqelsmmfekvqycrdlrdkyqtvslqkdgdnpkddkthwki ypeppppswhetekrfrgsskkehqkkdptmdefkfedceipgpndmvfkrdptcvyqvyedesslnenkpfvaipsirdyymdledliv assdgpaksfafrrlqyleakwnlyyllneytettesktnphrdfynvrkvdthvhhsacmnqkhllrfikykmkncpdevvihrdgreltlsq vfeslnltaydlsidtldmhahkdsfhrfdkfnlkynpvgesrlreiflktdnyiqgrylaeitkevfqdlenskyqmaeyrisiygrskdewdkl aawvldnklfspnvrwliqvprlydiykkaglvntfadivqnvfeplfevtkdpsthpklhvflqrvvgfdsvddeskldrrfhrkfptaaywd saqnppysywqyylyanmasintwrqrlgyntfelrphageagdpehllctylvaqginhgillrkvpfiqylyyldqipiamspvsnnalflt fdknpfysyfkrglnyslssddplqfaytkealieeysvaaliyklsnvdmcelarnsvlqsgferiikehwigenyeihgpegntiqktnvpnv rlafrdetlthelalvdkytnleeferlhg YALI0D16753g DNA: (SEQ ID NO: 138) atgttccgaacccgagttaccggctccaccctgcgatccttctccacctccgctgcccgacagcacaaggttgtcgtccttggcgccaacggaggc attggccagcccctgtctctgctgctcaagctcaacaagaacgtgaccgacctcggtctgtacgatctgcgaggcgcccccggcgttgctgccgat gtctcccacatccccaccaactccaccgtggccggctactctcccgacaacaacggcattgccgaggccctcaagggcgccaagctggtgctgat ccccgccggtgtcccccgaaagcccggcatgacccgagacgatctgttcaacaccaacgcctccattgtgcgagacctggccaaggccgtcggt gagcacgcccccgacgcctttgtcggagtcattgctaaccccgtcaactccaccgtccccattgtcgccgaggtgctcaagtccaagggcaagtac gaccccaagaagctcttcggtgtcaccaccctcgacgtcatccgagccgagcgattcgtctcccagctcgagcacaccaaccccaccaaggagta cttccccgttgttggcggccactccggtgtcaccattgtccccctcgtgtcccagtccgaccaccccgacattgccggtgaggctcgagacaagctt gtccaccgaatccagtttggcggtgacgaggttgtcaaggccaaggacggtgccggatccgccaccctttccatggcccaggctgccgcccgatt cgccgactctctcctccgaggtgtcaacggcgagaaggacgttgttgagcccactttcgtcgactctcctctgttcaagggtgagggcatcgacttct tctccaccaaggtcactcttggccctaacggtgttgaggagatccaccccatcggaaaggtcaacgagtacgaggagaagctcatcgaggctgcc aaggccgatctcaagaagaacattgagaagggtgtcaactttgtcaagcagaacccttaa Protein: (SEQ ID NO: 139) mfrtrvtgstlrsfstsaarqhkvvvlganggigqplslllklnknvtdlglydlrgapgvaadvshiptnstvagyspdnngiaealkgaklvlip agvprkpgmtrddlfntnasivrdlakavgehapdafvgvianpvnstvpivaevlkskgkydpkklfgvttldviraerfvsqlehtnptkey fpvvgghsgvtivplvsqsdhpdiageardklvhriqfggdevvkakdgagsatlsmaqaaarfadsllrgvngekdvveptfvdsplfkgeg idffstkvtlgpngveeihpigkvneyeeklieaakadlkkniekgvnfvkqnp YALI0D16247g DNA: (SEQ ID NO: 140) atgacacaaacgcacaatctgttttcgccaatcaaagtgggctcttcggagctccagaaccggatcgttctcgcacccttgactcgaaccagagctct gcccggaaacgtgccctcggatcttgccacagagtactacgcacaaagagcagcatctccaggcactctcctcatcaccgaggccacatacatctc ccccggatctgctggagtgcccattccaggagacggaatcgttccgggcatctggagtgacgagcagctcgaagcatggaaaaaggtgttcaag gccgtgcacgaccgaggatccaaaatctacgtccagctgtgggacattggacgtgtcgcatggtaccacaagctgcaggaactgggcaactactt ccctacaggcccctcagctatccccatgaagggagaggagagcgagcatctcaaggctctgactcactgggagatcaagggcaaggtggccctc tacgtcaacgctgccaagaacgccattgccgcaggcgctgatggcgtcgagatccactcggccaacggctaccttcccgacacatttctgagaag cgcctccaaccaacgaacagacgaatatggaggaagcatcgagaaccgggcccgattctcgctggagattgtcgacgctatcaccgaggccatt ggagcagacaaaaccgccatccgtctgtctccctggtccactttccaggacattgaggtgaatgacaccgagacccccgcacagttcacatacctg tttgagcagctgcagaagcgagccgacgagggaaagcagctggcctacgtgcatgtagttgagccccgactgtttggtccccccgagccctggg ccaccaatgagcctttcagaaaaatttggaagggtaacttcattagagcaggtggatacgatagagagactgctcttgaggatgcagacaagtcaga caacaccctgattgcctttggtcgagacttcattgccaatcctgatctcgtccaacgcctcaagaataacgagcctttggccaagtacgacagaacaa ccttctacgttccaggtgccaagggctacactgattaccctgcgtacaagatgtaa Protein: (SEQ ID NO: 141) mtqthnlfspikvgsselqnrivlapltrtralpgnvpsdlateyyaqraaspgtlliteatyispgsagvpipgdgivpgiwsdeqleawkkvfk avhdrgskiyvqlwdigrvawyhklqelgnyfptgpsaipmkgeesehlkalthweikgkvalyvnaaknaiaagadgveihsangylpdt flrsasnqrtdeyggsienrarfsleivdaiteaigadktairlspwstfqdievndtetpaqftylfeqlqkradegkqlayvhvveprlfgppep watnepfrkiwkgnfiraggydretaledadksdntliafgrdfianpdlvqrlknneplakydrttfyvpgakgytdypaykm YALI0A15972g DNA: (SEQ ID NO: 142) atggaagccaaccccgaagtccagaccgatatcatcacgctgacccggttcattctgcaggaacagaacaaggtgggcgcgtcgtccgcaatccc caccggagacttcactctgctgctcaactcgctgcagtttgccttcaagttcattgcccacaacatccgacgatcgaccctggtcaacctgattggcct gtcgggaaccgccaactccaccggcgacgaccagaagaagctggacgtgatcggagacgagatcttcatcaacgccatgaaggcctccggtaa ggtcaagctggtggtgtccgaggagcaggaggacctcattgtgtttgagggcgacggccgatacgccgtggtctgcgaccccatcgacggatcct ccaacctcgacgccggcgtctccgtcggcaccattttcggcgtctacaagctccccgagggctcctccggatccatcaaggacgtgctccgaccc ggaaaggagatggttgccgccggctacaccatgtacggtgcctccgccaacctggtgctgtccaccggaaacggctgcaacggcttcactctcga tgaccctctgggagagttcatcctgacccaccccgatctcaagctccccgatctgcgatccatctactccgtcaacgagggtaactcctccctgtggt ccgacaacgtcaaggactacttcaaggccctcaagttccccgaggacggctccaagccctactcggcccgatacattggctccatggtcgccgac gtgcaccgaaccattctctacggaggtatgtttgcctaccccgccgactccaagtccaagaagggcaagctccgacttttgtacgagggtttccccat ggcctacatcattgagcaggccggcggtcttgccatcaacgacaacggcgagcgaatcctcgatctggtccccaccgagatccacgagcgatcc ggcgtctggctgggctccaagggcgagattgagaaggccaagaagtaccttctgaaatga Protein: (SEQ ID NO: 143) meanpevqtdiitltrfilqeqnkvgassaiptgdftlllnslqfafkfiahnirrstlvnliglsgtanstgddqkkldvigdetttnamkasgkvkl vvseeqedlivfegdgryavvcdpidgssnldagvsvgtifgvyklpegssgsikdvlrpgkemvaagytmygasanlvlstgngcngftld dplgefilthpdlklpdlrsiysvnegnsslwsdnvkdyfkalkfpedgskpysaryigsmvadvhrtilyggmfaypadskskkgklrllye gfpmayiieqagglaindngerildlvpteihersgvwlgskgeiekakkyllk YALI0E11099g DNA: (SEQ ID NO: 144) atgcgactcactctgccccgacttaacgccgcctacattgtaggagccgcccgaactcctgtcggcaagttcaacggagccctcaagtccgtgtctg ccattgacctcggtatcaccgctgccaaggccgctgtccagcgatccaaggtccccgccgaccagattgacgagtttctgtttggccaggtgctgac cgccaactccggccaggcccccgcccgacaggtggttatcaagggtggtttccccgagtccgtcgaggccaccaccatcaacaaggtgtgctctt ccggcctcaagaccgtggctctggctgcccaggccatcaaggccggcgaccgaaacgttatcgtggccggtggaatggagtccatgtccaacac cccctactactccggtcgaggtcttgttttcggcaaccagaagctcgaggactccatcgtcaaggacggtctctgggacccctacaacaacatccac atgggcaactgctgcgagaacaccaacaagcgagacggcatcacccgagagcagcaggacgagtacgccatcgagtcctaccgacgggccaa cgagtccatcaagaacggcgccttcaaggatgagattgtccccgttgagatcaagacccgaaagggcaccgtgactgtctccgaggacgaggag cccaagggagccaacgccgagaagctcaagggcctcaagcctgtctttgacaagcagggctccgtcactgccggtaacgcctcccccatcaacg atggtgcttctgccgttgtcgttgcctctggcaccaaggccaaggagctcggtacccccgtgctcgccaagattgtctcttacgcagacgccgccac cgcccccattgactttaccattgctccctctctggccattcccgccgccctcaagaaggctggccttaccaaggacgacattgccctctgggagatca acgaggccttctccggtgtcgctctcgccaacctcatgcgactcggaattgacaagtccaaggtcaacgtcaagggtggagctgttgctctcggcc accccattggtgcctccggtaaccgaatctttgtgactttggtcaacgccctcaaggagggcgagtacggagttgccgccatctgcaacggtggag gagcttccaccgccatcgtcatcaagaaggtctcttctgtcgagtag Protein: (SEQ ID NO: 145) mrltlprlnaayivgaartpvgkfngalksvsaidlgitaakaavqrskvpadqideflfgqvltansgqaparqvvikggfpesveattinkvcs sglktvalaaqaikagdrnvivaggmesmsntpyysgrglvfgnqkledsivkdglwdpynnihmgnccentnkrdgitreqqdeyaiesy rranesikngafkdeivpveiktrkgtvtvsedeepkganaeklkglkpvfdkqgsvtagnaspindgasavvvasgtkakelgtpvlakivsy adaatapidftiapslaipaalkkagltkddialweineafsgvalanlmrlgidkskvnvkggavalghpigasgnrifvtlvnalkegeygva aicngggastaivikkvssve YALI0E34793g DNA: (SEQ ID NO: 146) atgtctgccaacgagaacatctcccgattcgacgcccctgtgggcaaggagcaccccgcctacgagctcttccataaccacacacgatctttcgtct atggtctccagcctcgagcctgccagggtatgctggacttcgacttcatctgtaagcgagagaacccctccgtggccggtgtcatctatcccttcggc ggccagttcgtcaccaagatgtactggggcaccaaggagactcttctccctgtctaccagcaggtcgagaaggccgctgccaagcaccccgaggt cgatgtcgtggtcaactttgcctcctctcgatccgtctactcctctaccatggagctgctcgagtacccccagttccgaaccatcgccattattgccgag ggtgtccccgagcgacgagcccgagagatcctccacaaggcccagaagaagggtgtgaccatcattggtcccgctaccgtcggaggtatcaagc ccggttgcttcaaggttggaaacaccggaggtatgatggacaacattgtcgcctccaagctctaccgacccggctccgttgcctacgtctccaagtc cggaggaatgtccaacgagctgaacaacattatctctcacaccaccgacggtgtctacgagggtattgctattggtggtgaccgataccctggtact accttcattgaccatatcctgcgatacgaggccgaccccaagtgtaagatcatcgtcctccttggtgaggttggtggtgttgaggagtaccgagtcat cgaggctgttaagaacggccagatcaagaagcccatcgtcgcttgggccattggtacttgtgcctccatgttcaagactgaggttcagttcggccac gccggctccatggccaactccgacctggagactgccaaggctaagaacgccgccatgaagtctgctggcttctacgtccccgataccttcgagga catgcccgaggtccttgccgagctctacgagaagatggtcgccaagggcgagctgtctcgaatctctgagcctgaggtccccaagatccccattga ctactcttgggcccaggagcttggtcttatccgaaagcccgctgctttcatctccactatttccgatgaccgaggccaggagcttctgtacgctggcat gcccatttccgaggttttcaaggaggacattggtatcggcggtgtcatgtctctgctgtggttccgacgacgactccccgactacgcctccaagtttctt gagatggttctcatgcttactgctgaccacggtcccgccgtatccggtgccatgaacaccattatcaccacccgagctggtaaggatctcatttcttcc ctggttgctggtctcctgaccattggtacccgattcggaggtgctcttgacggtgctgccaccgagttcaccactgcctacgacaagggtctgtcccc ccgacagttcgttgataccatgcgaaagcagaacaagctgattcctggtattggccatcgagtcaagtctcgaaacaaccccgatttccgagtcgag cttgtcaaggactttgttaagaagaacttcccctccacccagctgctcgactacgcccttgctgtcgaggaggtcaccacctccaagaaggacaacc tgattctgaacgttgacggtgctattgctgtttcttttgtcgatctcatgcgatcttgcggtgcctttactgtggaggagactgaggactacctcaagaac ggtgttctcaacggtctgttcgttctcggtcgatccattggtctcattgcccaccatctcgatcagaagcgactcaagaccggtctgtaccgacatcctt gggacgatatcacctacctggttggccaggaggctatccagaagaagcgagtcgagatcagcgccggcgacgtttccaaggccaagactcgatc atag Protein: (SEQ ID NO: 147) msanenisrfdapvgkehpayelfhnhtrsfvyglqpracqgmldfdfickrenpsvagviypfggqfvtkmywgtketllpvyqqvekaa akhpevdvvvnfassrsvysstmelleypqfrtiaiiaegvperrareilhkaqkkgvtiigpatvggikpgcfkvgntggmmdnivasklyr pgsvayvsksggmsnelnniishttdgvyegiaiggdrypgttfidhilryeadpkckiivllgevggveeyrvieavkingqikkpivawaigt casmfktevqfghagsmansdletakaknaamksagfyvpdtfedmpevlaelyekmvakgelsrisepevpkipidyswaqelglirkp aafistisddrgqellyagmpisevfkedigiggvmsllwfrrrlpdyaskflemvlmltadhgpavsgamntiittragkdlisslvaglltigtrf ggaldgaatefttaydkglsprqfvdtmrkqnklipgighrvksrnnpdfrvelvkdfvkknfpstqlldyalaveevttskkdnlilnvdgaia vsfvdlmrscgaftveetedylkngvlnglfvlgrsigliahhldqkrlktglyrhpwdditylvgqeaiqkkrveisagdvskaktrs YALI0D24431g DNA: (SEQ ID NO: 148) atgtcagcgaaatccattcacgaggccgacggcaaggccctgctcgcacactttctgtccaaggcgcccgtgtgggccgagcagcagcccatca acacgtttgaaatgggcacacccaagctggcgtctctgacgttcgaggacggcgtggcccccgagcagatcttcgccgccgctgaaaagacctac ccctggctgctggagtccggcgccaagtttgtggccaagcccgaccagctcatcaagcgacgaggcaaggccggcctgctggtactcaacaagt cgtgggaggagtgcaagccctggatcgccgagcgggccgccaagcccatcaacgtggagggcattgacggagtgctgcgaacgttcctggtcg agccctttgtgccccacgaccagaagcacgagtactacatcaacatccactccgtgcgagagggcgactggatcctcttctaccacgagggagga gtcgacgtcggcgacgtggacgccaaggccgccaagatcctcatccccgttgacattgagaacgagtacccctccaacgccacgctcaccaagg agctgctggcacacgtgcccgaggaccagcaccagaccctgctcgacttcatcaaccggctctacgccgtctacgtcgatctgcagtttacgtatct ggagatcaaccccctggtcgtgatccccaccgcccagggcgtcgaggtccactacctggatcttgccggcaagctcgaccagaccgcagagtttg agtgcggccccaagtgggctgctgcgcggtcccccgccgctctgggccaggtcgtcaccattgacgccggctccaccaaggtgtccatcgacgc cggccccgccatggtcttccccgctcctttcggtcgagagctgtccaaggaggaggcgtacattgcggagctcgattccaagaccggagcttctct gaagctgactgttctcaatgccaagggccgaatctggacccttgtggctggtggaggagcctccgtcgtctacgccgacgccattgcgtctgccgg ctttgctgacgagctcgccaactacggcgagtactctggcgctcccaacgagacccagacctacgagtacgccaaaaccgtactggatctcatgac ccggggcgacgctcaccccgagggcaaggtactgttcattggcggaggaatcgccaacttcacccaggttggatccaccttcaagggcatcatcc gggccttccgggactaccagtcttctctgcacaaccacaaggtgaagatttacgtgcgacgaggcggtcccaactggcaggagggtctgcggttg atcaagtcggctggcgacgagctgaatctgcccatggagatttacggccccgacatgcacgtgtcgggtattgttcctttggctctgcttggaaagcg gcccaagaatgtcaagccttttggcaccggaccttctactgaggcttccactcctctcggagtttaa Protein: (SEQ ID NO: 149) Msaksiheadgkallahflskapvwaeqqpintfemgtpklasltfedgvapeqifaaaektypwllesgakfvakpdqlikrrgkagllvlnk sweeckpwiaeraakpinvegidgvlrtflvepfvphdqkheyyinihsvregdwilfyheggvdvgdvdakaakilipvdieneypsnatl tkellahvpedqhqtlldfinrlyavyvdlqftyleinplvviptaqgvevhyldlagkldqtaefecgpkwaaarspaalgqvvtidagstkvsi dagpamvfpapfgrelskeeayiaeldsktgaslkltvlnakgriwtlvagggasvvyadaiasagfadelanygeysgapnetqtyeyaktvl dlmtrgdahpegkvlfigggianftqvgstfkgiirafrdyqsslhnhkvkiyvrrggpnwqeglrliksagdelnlpmeiygpdmhvsgivp lallgkrpknvkpfgtgpsteastplgv YALI0E14190g DNA: (SEQ ID NO: 150) atggttattatgtgtgtgggacctcagcacacgcatcatcccaacacagggtgcagtatatatagacagacgtgttccttcgcaccgttcttcacatatc aaaacactaacaaattcaaaagtgagtatcatggtgggagtcaattgattgctcggggagttgaacaggcaacaatggcatgcacagggccagtga aggcagactgcagtcgctgcacatggatcgtggttctgaggcgttgctatcaaaagggtcaattacctcacgaaacacagctggatgttgtgcaatc gtcaattgaaaaacccgacacaatgcaagatctctttgcgcgcattgccatcgctgttgccatcgctgtcgccatcgccaatgccgctgcggattatta tccctaccttgttccccgcttccgcacaaccggcgatgtctttgtatcatgaactctcgaaactaactcagtggttaaagctgtcgttgccggagccgct ggtggtattggccagcccctttctcttctcctcaaactctctccttacgtgaccgagcttgctctctacgatgtcgtcaactcccccggtgttgccgctga cctctcccacatctccaccaaggctaaggtcactggctacctccccaaggatgacggtctcaagaacgctctgaccggcgccaacattgtcgttatc cccgccggtatcccccgaaagcccggtatgacccgagacgatctgttcaagatcaacgctggtatcgtccgagatctcgtcaccggtgtcgcccag tacgcccctgacgcctttgtgctcatcatctccaaccccgtcaactctaccgtccctattgctgccgaggtcctcaagaagcacaacgtcttcaaccct aagaagctcttcggtgtcaccacccttgacgttgtccgagcccagaccttcaccgccgctgttgttggcgagtctgaccccaccaagctcaacatcc ccgtcgttggtggccactccggagacaccattgtccctctcctgtctctgaccaagcctaaggtcgagatccccgccgacaagctcgacgacctcgt caagcgaatccagtttggtggtgacgaggttgtccaggctaaggacggtcttggatccgctaccctctccatggcccaggctggtttccgatttgccg aggctgtcctcaagggtgccgctggtgagaagggcatcatcgagcccgcctacatctaccttgacggtattgatggcacctccgacatcaagcgag aggtcggtgtcgccttcttctctgtccctgtcgagttcggccctgagggtgccgctaaggcttacaacatccttcccgaggccaacgactacgagaa gaagcttctcaaggtctccatcgacggtctttacggcaacattgccaagggcgaggagttcattgttaaccctcctcctgccaagtaa Protein: (SEQ ID NO: 152) vvkavvagaaggigqplslllklspyvtelalydvvnspgvaadlshistkakvtgylpkddglknaltganivvipagiprkpgmtrddlfkin agivrdlvtgvaqyapdafvliisnpvnstvpiaaevlkkhnvfnpkklfgvttldvvraqtftaavvgesdptklnipvvgghsgdtivpllsltk pkveipadklddlvkriqfggdevvqakdglgsatlsmaqagfrfaeavlkgaagekgiiepayiyldgidgtsdikrevgvaffsvpvefgpe gaakaynilpeandyekkllkvsidglygniakgeefivnpppak YALI0E22649g DNA: (SEQ ID NO: 151) atgactggcaccttacccaagttcggcgacggaaccaccattgtggttcttggagcctccggcgacctcgctaagaagaagaccgtgagtattgaa ccagactgaggtcaattgaagagtaggagagtctgagaacattcgacggacctgattgtgctctggaccactcaattgactcgttgagagccccaat gggtcttggctagccgagtcgttgacttgttgacttgttgagcccagaacccccaacttttgccaccatacaccgccatcaccatgacacccagatgt gcgtgcgtatgtgagagtcaattgttccgtggcaaggcacagcttattccaccgtgttccttgcacaggtggtctttacgctctcccactctatccgagc aataaaagcggaaaaacagcagcaagtcccaacagacttctgctccgaataaggcgtctagcaagtgtgcccaaaactcaattcaaaaatgtcaga aacctgatatcaacccgtcttcaaaagctaaccccagttccccgccctcttcggcctttaccgaaacggcctgctgcccaaaaatgttgaaatcatcg gctacgcacggtcgaaaatgactcaggaggagtaccacgagcgaatcagccactacttcaagacccccgacgaccagtccaaggagcaggcca agaagttccttgagaacacctgctacgtccagggcccttacgacggtgccgagggctaccagcgactgaatgaaaagattgaggagtttgagaag aagaagcccgagccccactaccgtcttttctacctggctctgccccccagcgtcttccttgaggctgccaacggtctgaagaagtatgtctaccccg gcgagggcaaggcccgaatcatcatcgagaagccctttggccacgacctggcctcgtcacgagagctccaggacggccttgctcctctctggaa ggagtctgagatcttccgaatcgaccactacctcggaaaggagatggtcaagaacctcaacattctgcgatttggcaaccagttcctgtccgccgtgt gggacaagaacaccatttccaacgtccagatctccttcaaggagccctttggcactgagggccgaggtggatacttcaacgacattggaatcatcc gagacgttattcagaaccatctgttgcaggttctgtccattctagccatggagcgacccgtcactttcggcgccgaggacattcgagatgagaaggtc aaggtgctccgatgtgtcgacattctcaacattgacgacgtcattctcggccagtacggcccctctgaagacggaaagaagcccggatacaccgat gacgatggcgttcccgatgactcccgagctgtgacctttgctgctctccatctccagatccacaacgacagatgggagggtgttcctttcatcctccg agccggtaaggctctggacgagggcaaggtcgagatccgagtgcagttccgagacgtgaccaagggcgttgtggaccatctgcctcgaaatgag ctcgtcatccgaatccagccctccgagtccatctacatgaagatgaactccaagctgcctggccttactgccaagaacattgtcaccgacctggatct gacctacaaccgacgatactcggacgtgcgaatccctgaggcttacgagtctctcattctggactgcctcaagggtgaccacaccaactttgtgcga aacgacgagctggacatttcctggaagattttcaccgatctgctgcacaagattgacgaggacaagagcattgtgcccgagaagtacgcctacggc tctcgtggccccgagcgactcaagcagtggctccgagaccgaggctacgtgcgaaacggcaccgagctgtaccaatggcctgtcaccaagggct cctcgtga Protein: (SEQ ID NO: 153) mtgtlpkfgdgttivvlgasgdlakkktfpalfglyrngllpknveiigyarskmtqeeyherishyfktpddqskeqakkflentcyvqgpyd gaegyqrlnekieefekkkpephyrlfylalppsvfleaanglkkyvypgegkariiiekpfghdlassrelqdglaplwkeseifridhylgke mvknlnilrfgnqflsavwdkntisnvqisfkepfgtegrggyfndigiirdviqnhllqvlsilamerpvtfgaedirdekvkvlrcvdilnidd vilgqygpsedgkkpgytdddgvpddsravtfaalhlqihndrwegvpfilragkaldegkveirvqfrdvtkgvvdhlprnelviriqpsesi ymkmnsklpgltaknivtdldltynrrysdvripeayeslildclkgdhtnfvrndeldiswkiftdllhkidedksivpekyaygsrgperlkq wlrdrgyvrngtelyqwpvtkgss YALI0B15598g DNA: (SEQ ID NO: 154) atgactgacacttcaaacatcaagtgagtattgccgcacacaattgcaatcaccgccgggctctacctcctcagctctcgacgtcaatgggccagca gccgccatttgaccccaattacactggttgtgtaaaaccctcaaccacaatcgcttatgctcaccacagactacgacttaaccaagtcatgtcacaggt caaagtaaagtcagcgcaacaccccctcaatctcaacacacttttgctaactcaggcctgtcgctgacattgccctcatcggtctcgccgtcatgggc cagaacctgatcctcaacatggccgaccacggtaagtatcaattgactcaagacgcaccagcaagatacagagcatacccagcaatcgctcctctg ataatcgccattgtaacactacgttggttagattgatctaaggtcgttgctggttccatgcacttccacttgctcatatgaagggagtcaaactctattttg atagtgtcctctcccatccccgaaatgtcgcattgttgctaacaataggctacgaggttgttgcctacaaccgaaccacctccaaggtcgaccacttcc tcgagaacgaggccaagggtgagtatccgtccagctatgctgtttacagccattgaccccaccttcccccacaattgctacgtcaccattaaaaaaca aaattaccggtatcggcaagctagactttcatgcaacctacgcagggtaacaagttgagtttcagccgtgcaccttacaggaaaaccagtcatacgc cgaggcagtgtgaaagcgaaagcacacagcctacggtgattgattgcatttttttgacataggagggaaacacgtgacatggcaagtgcccaacac gaatactaacaaacaggaaagtccattattggtgctcactctatcaaggagctgtgtgctctgctgaagcgaccccgacgaatcattctgctcgttaag gccggtgctgctgtcgattctttcatcgaacagctcctgccctatctcgataagggtgatatcatcattgacggtggtaactcccacttccccgactcca accgacgatacgaggagcttaacgagaagggaatcctctttgttggttccggtgtttccggcggtgaggagggtgcccgatacggtccctccatcat gcccggtggaaacaaggaggcctggccccacattaagaagattttccaggacatctctgctaaggctgatggtgagccctgctgtgactgggtcgg tgacgctggtgccggccactttgtcaagatggttcacaacggtattgagtatggtgacatgcagcttatctgcgaggcttacgacctcatgaagcgag gtgctggtttcaccaatgaggagattggagacgttttcgccaagtggaacaacggtatcctcgactccttcctcattgagatcacccgagacatcttca agtacgacgacggctctggaactcctctcgttgagaagatctccgacactgctggccagaagggtactggaaagtggaccgctatcaacgctcttg accttggtatgcccgtcaccctgatcggtgaggccgtcttcgctcgatgcattctgccctcaagcaggagcgtgtccgagcttccaaggttcttgatg gccccgagcccgtcaagttcactggtgacaagaaggagtttgtcgaccagctcgagcaggccctttacgcctccaagatcatctcttacgcccagg gtttcatgcttatccgagaggccgccaagacctacggctgggagctcaacaacgccggtattgccctcatgtggcgaggtggttgcatcatccgatc cgtcttccttgctgacatcaccaaggcttaccgacaggaccccaacctcgagaacctgctgttcaacgacttcttcaagaacgccatctccaaggcc aacccctcttggcgagctaccgtggccaaggctgtcacctggggtgttcccactcccgcctttgcctcggctctggctttctacgacggttaccgatct gccaagctccccgctaacctgctccaggcccagcgagactacttcggcgcccacacctaccagctcctcgatggtgatggaaagtggatccacac caactggaccggccgaggtggtgaggtttcttcttccacttacgatgcttaa Protein: (SEQ ID NO: 155) mtdtsnikpvadialiglavmgqnlilnmadhgyevvaynrttskvdhfleneakgksiigahsikelcallkrprriillvkagaavdsfieqll pyldkgdiiidggnshfpdsnrryeelnekgilfvgsgvsggeegarygpsimpggnkeawphikkifqdisakadgepccdwvgdagag hfvkmvhngieygdmqliceaydlmkrgagftneeigdvfakwnngildsflieitrdifkyddgsgtplvekisdtagqkgtgkwtainald lgmpvtligeavfarclsalkqervraskvldgpepvkftgdkkefvdqleqalyaskiisyaqgfmlireaaktygwelnnagialmwrggci irsvfladitkayrqdpnlenllfndffknaiskanpswratvakavtwgvptpafasalafydgyrsaklpanllqaqrdyfgahtyqlldgdgk wihtnwtgrggevssstyda YALI0D06303g DNA: (SEQ ID NO: 156) atgctcaaccttagaaccgcccttcgagctgtgcgacccgtcactctggtgagtatctcggagcccgggacggctaccaacacacaagcaagatg caacagaaaccggactttttaaatgcggattgcggaaaatttgcatggcggcaacgactcggagaaggagcgggacaattgcaatggcaggatgc cattgacgaactgagggtgatgagagaccgggcctccgatgacgtggtggtgacgacagcccggctggtgttgccgggactgtctctgaaaagcaatt tctctatctccggtctcaacagactccccttctctagctcaattggcattgtcttcagaaggtgtcttagtggtatccccattgttatcttcttttcccca atgtcaatgtcaatgtcaatggctccgacctctttcacattaacacggcgcaaacacagataccacggaaccgactcaaacaaatccaaagagacg cagcggaataattggcatcaacgaacgatttgggatactctggcgagaatgccgaaatatttcgcttgtcttgttgtttctcttgagtgagttgtttgtgaa gtcgtttggaagaaggttcccaatgtcacaaaccataccaactcgttacagccagcttgtaatcccccacctcttcaatacatactaacgcagacccg atcctacgccacttccgtggcctctttcaccggccagaagaactccaacggcaagtacactgtgtctctgattgagggagacggtatcggaaccga gatctccaaggctgtcaaggacatctaccatgccgccaaggtccccatcgactgggaggttgtcgacgtcacccccactctggtcaacggcaaga ccaccatccccgacagcgccattgagtccatcaaccgaaacaaggttgccctcaagggtcccctcgccacccccatcggtaagggccacgtttcc atgaacctgactctgcgacgaaccttcaacctgttcgccaacgtccgaccttgcaagtccgtcgtgggctacaagaccccttacgagaacgtcgac accctgctcatccgagagaacactgagggtgagtactccggtatcgagcacaccgtcgtccccggtgtcgttcagtccatcaagctgatcacccga gaggcttccgagcgagtcatccggtacgcttacgagtacgccctgtcccgaggcatgaagaaggtccttgttgtccacaaggcctctattatgaagg tctccgatggtcttttccttgaggttgctcgagagctcgccaaggagtacccctccattgacctttccgtcgagctgatcgacaacacctgtctgcgaat ggtccaggaccccgctctctaccgagatgtcgtcatggtcatgcccaacctttacggtgacattctgtccgatcttgcctccggtcttatcggtggtctt ggtctgaccccctccggtaacatgggtgacgaggtctccatcttcgaggccgtccacggatccgctcccgacattgctggcaagggtcttgctaac cccactgctctgctgctctcctccgtgatgatgctgcgacacatgggtctcaacgacaacgccaccaacatcgagcaggccgtctttggcaccattg cttccggccccgagaaccgaaccaaggatcttaagggtaccgccaccacttctcactttgctgagcagattatcaagcgactcaagtag Protein: (SEQ ID NO: 157) mlnlrtalravrpvtltrsyatsvasftgqknsngkytvsliegdgigteiskavkdiyhaakvpidwevvdvtptlvngkttipdsaiesinrnkv alkgplatpigkghvsmnltlrrtfnlfanvrpcksvvgyktpyenvdtllirentegeysgiehtvvpgvvqsiklitreaserviryayeyalsrg mkkvlvvhkasimkvsdglflevarelakeypsidlsvelidntclrmvqdpalyrdvvmvmpnlygdilsdlasgligglgltpsgnmgde vsifeavhgsapdiagkglanptalllssvmmlrhmglndnatnieqavfgtiasgpenrtkdlkgtattshfaeqiikrlk

Example 13 Determination of Lipid Levels of Y. Lipolytica

12A. Determination of Lipid Levels of Y. Lipolytica in Various Growth Conditions of Varying Carbon to Nitrogen Ratios.

Shake flask testing was conducted using carbon to nitrogen (C/N) ratios of 160, 80, 60, 40, 30, 20, and 10 with yeast nitrogen base being the base medium providing vitamins, trace elements and salts Ammonium sulfate (which contains 21% nitrogen) was used as the nitrogen source and glucose (which contains 40% carbon) was used as the carbon source at a concentration of 30 g/L. The concentrations of ammonium sulfate corresponding to these ratios are: 0.36, 0.71, 0.95, 1.43, 1.91, 2.86, and 4.6 g/L, respectively. Uracil was supplemented at 0.2 mM. As controls, strains were also grown in yeast extract-peptone with 50 g/L of glucose (media in which lipids do not accumulate at high levels) and yeast extract-peptone with 5% olive oil (v/v) (media in which lipids accumulate at high levels).

Strain MF760 (10-14 ml of culture) was harvested after 4 days of growth at 30° C., during which time the cultures were shaking at 250 rpm. Following harvesting, cells were washed three times with water, with the exception of the oil-grown cells which were washed three times in 0.5% BSA and one time with water before lipid extractions. Lipids were extracted as described in Folch J, Lees, M, and Stanley, G. H. S. J. Biol. Chem. 226: 497-509, 1957. In brief, cell pellets were resuspended in 6 ml of water. A 1 ml aliquot was transferred to a pre-weighed tube with a hole on the lid, spun down and the cell pellet lyophilized overnight to determine the dry cell weight. The remaining 5 ml were placed in a 15 ml Falcon tube and spun down. Cell pellets were frozen at −20° C. until extractions were performed.

Two to three volumes of a Zymolyase solution (2 mg/ml Zymolyase 100T in 1M Sorbitol, 50 mM EDTA and 0.01% β-mercaptoethanol) was added to each cell pellet and placed at 37° C. with constant agitation for 1 hr. Two volumes of cubic zirconia beads were added to each tube and vortexed for 15-20 min. Samples were viewed under a microscope to ensure cell breakage before continuing with extractions. After cell breakage was complete, 6 ml of extraction solvent was added (a 2:1 mix of chloroform and methanol) and mixed. The mixture was spun down for 5 min at 3000 rpm and the organic layer was transferred to a clean tube. NaCl was added to the remaining aqueous layer to make it a 0.29% NaCl solution. 6 ml of extraction solvent was added and mixed, and the mixture was spun down for 5 min. The organic layers were pooled and filtered through a 0.2 μm filter to get rid of any cell debris. The extract was washed with 0.2 volumes of 0.29% NaCl solution and another 6 ml of extraction solvent added and mixed. Mixtures were spun and the organic layer was placed in a pre-weighed glass vial, the solvent was evaporated under a flow on nitrogen and the vial was weighed again to determine the weight of the lipid extracted. The dry cell weight is used to determine the percentage of lipid per dry cell weight. The lipid accumulation results are in the Table 48 below:

TABLE 48 Lipid accumulation under various carbon: nitrogen ratio growth conditions C/N Ratio % lipid YNB 160 61 3% Glucose 80 49 60 34 40 17 30 16 20 14 10 15 YEP 5% Glucose 22 5% olive oil 38 Other nitrogen sources tested were proline (12% nitrogen), sodium glutamate (7% nitrogen), soy acid hydrolysate (12% nitrogen), and yeast extract-peptone (26.8% nitrogen). All nitrogen sources tested at C/N ratios of 80 (with glucose as a carbon source), had significantly larger lipid bodies than at C/N ratios of 10 (also with glucose as a carbon source).

Strains MF858 and MF921 (example 2F and 2H) were harvested after 4 days of growth at 30° C. (3% glucose was used as the carbon source). Cells were washed three times with water and lipids extracted as described above. Lipid accumulation data for soy hydrolysate, yeast extract-peptone and yeast nitrogen base, used as a control, are listed in Table 49 below.

TABLE 49 Lipid accumulation under different carbon and nitrogen conditions with various nitrogen sources % lipid C/N Ratio MF858 MF921 Soy hydrolysate 80 36 36 60 36 35 10 14 15 Yeast Extract- 80 37 37 Peptone 10 15 14 Yeast Nitrogen 80 37 38 Base 10 13 11 12b. Determination of Lipid Levels Under High Carbon and Phosphate or Magnesium Limiting Conditions.

To test whether other nutrient limitations, under high carbon conditions, will allow for higher lipid accumulation, phosphate or magnesium limiting conditions were tested. For phosphate limiting conditions, yeast nitrogen base medium without phosphate was prepared. Shake flask testing was performed using carbon to phosphate ratios ranging from 5376 down to 42. This range corresponds to 7.8 mg/L up to 1 g/L, respectively, and the latter concentration corresponds to that are commonly used in yeast nitrogen base medium. Glucose, at 30 g/L, was used at the carbon source. Potassium phosphate monobasic (containing 28.7% phosphate) was used as the phosphate source.

For magnesium limiting conditions, yeast nitrogen base medium without magnesium was prepared. Shake flask testing was conducted using carbon to magnesium ratios ranging from 31360 down to 245. This range corresponds to 0.375 mg/L up to 0.5 g/L, and the latter magnesium concentration corresponds to that commonly used in yeast nitrogen base. Glucose, at 30 g/L, was used as the carbon source. Magnesium sulfate (containing 9.8% magnesium) was used as the magnesium source.

Strains MF858 and MF921 were harvested after 4 days of growth at 30° C., during which time the cultures were shaking at 250 rpm. Cells were washed three times with water before lipid extraction. Lipids were extracted as described above. Lipid accumulation data is listed in the Table 50 below:

TABLE 50 Lipid accumulation in phosphate or magnesium limiting growth conditions % Lipid g/L MF858 MF921 phosphate 1 14 14 0.0625 18 20 0.0313 34 41 0.0156 62 63 0.0078 83 76 magnesium 0.5 12 11 0.0313 NA 16 0.0156 NA 25 0.0078 NA 42 0.0039 48 48

Example 14 Effect of Temperature on Carotenoid Production

MF740 was transformed with pMB4719 with SalI, and a Ura⁺ colony was designated ML878. MF740 was transformed with pMB4629 cleaved with SalI, an Ade⁺ colony was designated ML857, and subsequently transformed with pMB4719 cleaved with SalI, to create ML836. ML878 and ML836 were grown for 4 days in YPD at 20° C., 24° C., and 28° C., and carotenoids were extracted and analyzed by HPLC. β-carotene or zeaxanthin yield (% dry cell weight) at 20° C. was chosen as a standard against which yields at other temperatures were compared. In addition, the ratio of zeaxanthin/carotenoid (% dry cell weight) was calculated for each temperature. Whereas the β-carotene levels fell with decreasing temperatures, the ratio of zeaxanthin to β-carotene increased with lower temperatures (Table 51).

TABLE 51 Effect of Temperature on carotenoid production Temperature β-carotene Zeaxanthin Zeaxanthin/ (β-carotene ratio Strain (° C.) yield* yield** (% dry cell weight)*** MF878 20 1.0 1.0 2.4 MF878 24 1.3 1.0 1.9 MF878 28 3.2 1.5 1.1 MF836 20 1.0 1.0 1.0 MF836 24 1.9 1.2 0.7 MF836 28 3.4 1.1 0.4 *β-carotene yield is calculated as % DCW β-carotene at 20° C. divided by % DCW β-carotene at each temperature **Zeaxanthin yield is calculated as % DCW zeaxanthin at 20° C. divided by % DCW zeaxanthin at each temperature ***Zeaxanthin/β-carotene ratio is calculated as % DCW zeaxanthin divided by % DCW β-carotene

Example 15 Construction of a Recyclable URA3 Marker Plasmid pMB5082

To create a selectively excisable (“recyclable”) URA3 marker, an 860 bp SpeI-SacI (blunt ended with T4 DNA ligase) fragment (containing the URA3 promoter and the first 121 nucleotides of the URA3 gene) from plasmid pMB4691 was inserted into the SpeI-NotI sites of plasmid pMB4534 to create pMB5055.

The URA3 promoter was excised from pMB5055 as an 878 bp fragment by XbaI-SpeI digest, and was ligated into XbaI-cleaved pMB4691. Orientation of the promoter was verified by restriction digest. The resulting plasmid, designated pMB5082, contained the URA3 promoter both upstream of the URA3 gene and downstream of its terminator. This cassette, once integrated into the Yarrowia genome, permits excision of the URA3 marker by homologous recombination between the two copies of the URA3 promoter. Colonies containing the excision may be selected on 5-FOA.

Example 16 Effects of Mutations in the Transcriptional Regulator, SPT8 on Carotenoid Production

Y. Lipolytica strain ML1018 was isolated by plasmid insertion mutagenesis. ML1018 was darker in hue, shiny, exclusively yeast-form rather than partial mycelial morphology and exhibited increased carotenoid levels when compared to its sibling transformants. Sequence analysis identified the site of ML1018 plasmid insertion between base pairs 701 and 702 of the SPT8 coding sequence. Experiments were undertaken to examine carotenoid levels in a targeted SPT8 disruption strain.

A 2.5 kb fragment containing the SPT8 gene (YALI0E23804g) with its endogenous promoter and terminator was amplified from genomic DNA isolated from Y. lipolytica strain NRRL Y-1095 using primers: MO5651 (5′-CACAAACTAGTGTCAGGAATATGAAACCAGG-3′) (SEQ ID NO:158) and MO5652 (5′-CACAAACTAGTGCATGTGATAGGAAGGAGGA-3′) (SEQ ID NO:159). Plasmid pMB5083 was constructed by phosphorylating the 2.5 kb SPT8 fragment with T4 polynucleotide kinase and ligating the phosphorylated fragment with desphosphorylated, EcoRV-digested pBluescriptSK−.

A 3.4 kb fragment containing the TEF1 promoter, XPR terminator, and a recyclable URA3 marker was isolated from plasmid pMB5082 bp Acc65I and XbaI (subsequently made blunt with Klenow) digestion. This fragment was cloned into the BsiWI and SmaI sites of pMB5083 to create pMB5086. BamHI-XbaI digestion of pMB5086 yields a 5.6 kb Y. Lipolytica SPT8 disruption fragment containing the TEF1 promoter and XPR terminators followed by a recyclable URA3 marker between base pairs 752 and 753 of the SPT8 coding sequence (SPT8::URA3 disruption cassette).

A 3.6 kb fragment containing the XPR terminator and ADE1 gene was excised from plasmid pMB4629 by MluI and EcoRV digest and subsequently cloned into MluI-Pml1-digested pMB5086. The resulting plasmid, pMB5124, contains a 5.8 kb BamHI-XbaI SPT8 disruption cassette similar to that in pMB5086, with the distinction that the recyclable URA3 marker is replaced with a non-recyclable ADE1 marker (SPT8::ADE1 disruption cassette).

Y. lipolytica strains MF740 and MF746 (both ade1 ura3) are transformed with a 5.8 kb BamHI-XbaI fragment from pMB5124 (spt8::ADE1). spt8 disruptants are distinguished from ectopic integrants by colony morphology, as spt8 strains are shinier, darker in hue, and less mycelial than SPT8 strains. Correct integration may be assayed by PCR or by Southern blotting. Carotenoid yield is assayed in spt8 disrupted and SPT8⁺ strains by harvesting carotenoids after a four-day fermentation in YPD shake flasks at 30° C.

Example 17 Construction Plasmid pMB4844 Encoding a Chimeric β-Carotene Hydroxylase-Ketolase (crtZW)

A β-carotene hydroxylase:β-carotene ketolase chimera is constructed as follows. First, a 0.5 kb fragment containing crtZ from Erythrobacter litoralis is amplified from pMB4715, a plasmid containing a copy of the crtZ gene, using primers MO4814: 5′-CACAACGTCTCTCTAGACACAAAAATGAGCT-3′ (SEQ ID NO:160) and MO4816: 5′-CACAACGTCTCAGCCGGCACCTGCTCCCATAGAATCTCG-3′ (SEQ ID NO:161) and the resulting fragment is digested with XbaI and BsmBI. Similarly, a 0.8 kb fragment containing crtW from Parvularcula bermudensis is amplified from pMB4731, a plasmid containing a copy of the crtW gene, with primers MO5060: 5′-CACAAGAAGACAACGGCGCAGGAGCCATGGACCCTACCGGAGACG-3′ (SEQ ID NO:162) and MO5061: 5′-CACAAGAAGACAACGCGTTTAAGGGCCGGTTCTCTTTC-3′ (SEQ ID NO:163) and the resulting fragment is digested with BbsI and MluI. The digested fragments containing the crtZ and crtW genes are then ligated in a three-piece reaction into NheI-MluI cleaved vector pMB4691 to create pMB4844. Sequence analysis confirms the creation of an in-frame fusion of crtZ and crtW placed under control of the TEF1 promoter and the XPR terminator. The chimeric sequence is designated crtZW. The amino acid sequence of crtZW is:

(SEQ ID NO: 175) mswwaialivfgavvgmeffawfahkyimhgwgwswhrdhhephdntle kndlfavvfgsvaallfvigalwsdplwwaavgitlygviytlvhdglvh qrywrwtpkrgyakrlvqahrlhhatvgkeggvsfgfvfardpaklka elkqqreqglavvrdsmgagagagamdptgdvtasprpqttipvrq alwglslagaiiaawvfmhigfvffapldpivlalapviillqswls vglfiishdaihgslapgrpafnramgrlcmtlyagfdfdrmaaa hhrhhrspgtaadpdfsvdspdrplpwfgaffrryfgwrpfl tvnavvftywlvlganpvnivlfygvpallsagqlfyfgtflphrherq gfadhhrarsvrspymlslvtcyhfggyhhehhlfphepwwrlpq rggwerdrrkrtgp

Example 18 pH Effects on Total Carotenoid Yield and Hydroxylation of Beta-Carotene

The effect of altering pH on total carotenoid yield and relative amount of individual carotenoids was investigated. Strain ML1011 (MF740 transformed with multiple integrated copies of the X. autotrophicus crtZ gene) which accumulates a mixture of carotenoids comprising beta-carotene, beta-cryptoxanthin, and zeaxanthin was fermented under the following parameters.

-   -   Batch medium: YPD     -   Temperature setpoint: 30° C.     -   Initial volume: 210 ml     -   Vessel volume: 400 ml     -   Agitation rate: 1000 rpm     -   Feed: 40% glucose     -   Feed rate: 2 ml/hour, starting at 24th hour after inoculation

Four separate fermentor units were setup and the pH was controlled as follows:

-   -   Unit 1: pH 5.5     -   Unit 2: pH 7.0     -   Unit 3: pH 7.0 at inoculation, continuously rising to a setpoint         of pH 8.0 at 48 hours (change of 0.021 pH units/hour)     -   Unit 4: pH 7.0 at inoculation, continuously rising to a setpoint         of pH 9.0 at 48 hours (change of 0.042 pH units/hour

Additionally, the glucose feed of unit 4 was halted at 64 hours (see below). FIG. 12a depicts accumulation of total carotenoid (absorbance units per unit dry cell weight) throughout the fermentation. Excluding the last timepoint, units 1 and 2 accumulated similar amounts of carotenoid throughout the run. This result, consistent with previous experiments, suggests that varying the pH in the range of 5.5-7.0 does not affect total carotenoid yield. During this same period, fermentor unit 3 accumulated more carotenoid than units 1 and 2, suggesting setting pH to be within the range of 7.0-8.0 improves the rate of carotenoid biosynthesis. In unit 4, carbon dioxide evolution (indicating metabolic activity) and carotenoid accumulation started to fall precipitously when the unit reached pH 8.3 at approximately 31 hours (FIG. 12d ), suggesting toxicity due to high pH. The feed to this unit was therefore stopped at 64 hours. Together, these results suggest that carotenoid yield may be maximized by maintaining pH within the range 7.0-8.0, while pH levels below or above this range were ineffective or toxic, respectively.

FIG. 12b depicts accumulation of zeaxanthin (absorbance units per dry cell weight; AU) over the course of the fermentation. As seen in FIG. 12b , zeaxanthin accumulation improves with increasing pH. Unit 4, at highest pH, shows superior zeaxanthin accumulation compared to all other units until hour 64, when its feed was terminated. Likewise, unit 3, maintained at pH 8.0 after hour 48, shows significantly improved zeaxanthin accumulation over units 1 and 2, maintained at pH 5.5 and 7.0, respectively. These results indicate that hydroxylation of beta carotene to zeaxanthin is favored by higher pH.

FIG. 12c depicts the fraction of carotenoid as zeaxanthin (AU zeaxanthin/AU total carotenoid) throughout the course of the fermentation. Unit 3 hydroxylated a greater fraction of beta-carotene than units 1 and 2, in addition to producing more total carotenoid (FIG. 12a ) and more total zeaxanthin (FIG. 12b ). This result demonstrates that at pH 8.0, zeaxanthin accumulation outpaced the global increase in carotenoid biosynthesis also seen at this pH

As seen in FIG. 12e , biomass accumulated fastest in unit 3 and remained above all other units until the hundred-thirtieth or so hour of fermentation. This unit similarly was the most metabolically active, as shown by its increased rate of carbon dioxide evolution compared to the other units over the same time period (FIG. 12d ). The subsequent decline in biomass in this unit may be attributed to accelerated metabolism of carbon stored as intracellular oil, relative to the other three units. Thus, it appears that the pH range of 7.0-8.0 enables Yarrowia lipolytica both to accumulate biomass and metabolize stored carbon at rates faster than it is able at lower pH.

Together, these results indicate that total biomass accumulation, percentage of biomass representing carotenoid accumulation, and the hydroxylation of beta-carotene to zeaxanthin may be manipulated by maintaining fermentation pH in the approximate range of 7.0-8.0. Moreover, these results suggest that within this same range, an optimum pH may be selected at which to maximize production of both non-oxygenated carotenoids and xanthophylls (e.g. hydroxylation of b-carotene to zeaxanthin and total carotenoid production.

Example 19 Lycopene Epsilon Cyclase Sequences

The DNA and proteins they encode of the certain lycopene epsilon cyclase sequences are provided below. Corresponding Genbank Accession and GI numbers are found in table 23.

Ostreococcus lucimarinus sequence XP_001422490 DNA (SEQ ID NO: 164) ATGAAGGATGATCGCGAATGGATTGCGTTTCAACAGCGCAAGGTGTTTAGTGAGCAAAA GCAAATCAAAGAGTACCTCAGTGCTTTGAACGACCGCGACAAGGTCGACGTTCTCGTTGT CGGTGCGGGCCCCGCAGGTCTGGCGATCGCAGCGGAGACGGCGAAGAAGGGTCTTTCTG TTGGTCTCGTCGCACCAGACACCCCGTTCGTGAACAACTACGGAGTATGGCTCGACGAGT TCAAAGATCTAGGGCTCGAACACTGCTTGCTTCATAAGTATGACGACGCATTGGTTTGGT TCGATGATTCTGATCCTGCGAGTGGAACTGAACTCGGTCGACCTTACGGTCAAGTGTGCC GCAGGCGTCTTCGCGACCATTTGTTGAAGGAGTGCGCGGCGGCTGGCGTCAAGTATTTAC CAGGCCTGGTAGATTTTGTGCGTCACGGTGACGTCGAAAAGAACGAGTTAGCCGAAGCA AACAGAGGCCAGCAATTCACGTTGAATTCGCGTCTCGTCGTTGCCGGCACCGGTCACAAC CGCGACATGCTCAGCTACGAAGAGGGTGCGCCGCCGGGCTGGCAGACTGCGTATGGCGT TGAGGTGCGCATTCCGAACCACGGTTTTCCCGTGAACAAGGCCGTGTTCATGGATTTTCG TCAAAGCGATCCGGAGGCGATGAAAGAGGAACAAGACGAGGGCGTTTGGCGCGTGCCG TCTTTCCTTTACGTGTTACCCGTGGACAAGGATGTGGTGTTCGTCGAGGAGACGTGCCTC GTCGCGCGCGTACAAGTGCCGTTCGATGAACTCAAACGGCGATTGTATCGTCGTATGAAG CGGATGGGTATGGAAATCGTCGAAGAAGACATCTTGGAAGTCGAGGCGAGTTGGATTCC ACTGGGCGGTACCCCGCCGGTTGCCCCGCAACGCACCATCGCGTACGGTGCAGCAGCCG GCATGGTCCACCCTGCGTCTGGCTACTCCGTCGTAAACAGTATTAGCAAAGCTCCGCGTG TTGCGACGGCCATGGCCGAAGGCTTGAAGGAGGGTGGCGAGATTGAGGCGAGCCGAAG AGCGTGGGAAATCCTTTGGGGTGCGGAGCCACGAAGACAAATCGGTTTCTACCAGTTCG GTATGGAGCTTCTCATGTCGCTTCGCATCGAGCAGATGCGCAACTTCTTTAGTACCTTCTT TGCGCTTCCAACAAATCTGAGCAGAGGATTTTTGGGTAACAGATTGTCGAGCTCAGAGTT GATCATGTTTGCTCTCACTACGTTCGCAATTGGTAACAACGAACTTCGTGGGTTGTTGCTC GCTCACCTGGTTTCA Protein (SEQ ID NO: 165) MKDDREWIAFQQRKVFSEQKQIKEYLSALNDRDKVDVLVVGAGPAGLAIAAETAKKGLSVG LVAPDTPFVNNYGVWLDEFKDLGLEHCLLHKYDDALVWFDDSDPASGTELGRPYGQVCRR RLRDHLLKECAAAGVKYLPGLVDFVRHGDVEKNELAEANRGQQFTLNSRLVVAGTGHNRD MLSYEEGAPPGWQTAYGVEVRIPNHGFPVNKAVFMDFRQSDPEAMKEEQDEGVWRVPSFL YVLPVDKDVVFVEETCLVARVQVPFDELKRRLYRRMKRMGMEIVEEDILEVEASWIPLGGTP PVAPQRTIAYGAAAGMVHPASGYSVVNSISKAPRVATAMAEGLKEGGEIEASRRAWEILWG AEPRRQIGFYQFGMELLMSLRIEQMRNFFSTFFALPTNLSRGFLGNRLSSSELIMFALTTFAIGN NELRGLLLAHLVS lycopene epsilon cyclase (Diospyros kaki) sequence BAE94036 DNA (SEQ ID NO: 166) ACTACGGCGTATGGGAGGATGAATTTAGAGATCTTGGACTTGAAAGGTGTATTGAACAT GTTTGGAGAGACACAATTGTATATCTTGATGACAATGATCCCATTCTGATTGGTCGTGCT TATGGACGAGTTAGTCGTCACTTGCTCCACGAGGAGCTATTAAGAAGGTGTGTGGAGTCA GGTGTTTCATATTTGAGCTCAAAAGTGGAAAGAATTATTGAAACTACGAATGGGCAGAG TCTCATAGAGTGCGGAACTGATGTTGTTGTCCCATGCAGGCTTGCTACTGTTGCTTCGGG AGCAGCTTCTGGGAAACTTTTGAAGTTTGAGGTGGGAGGACCCAGAGTTTCTGTTCAAAC AGCTTATGGTGTGGAGGTTGAGGTGGAAAACAATCCATATGACCCCAACTTGATGGTTTT CATGGATTACAGAGACTATGCCAAACAAAAAGTTCAGCCTTTGGAAGCACAATATCCAA CATTTCTTTATGCCATGCCTATGTCCCCTACAAGAGTCTTCTTTGAGGAAACTTGTTTGGC TTCAAAGGATGCCATGCCTTTTGATCTATTAAAGAGGAAACTCATGGACAGATTAGAGAC AATGGGAGTCCATGTTCTAAAAACGTATGAGGAGGAATGGTCTT Protein (SEQ ID NO: 167) YGVWEDEFRDLGLERCIEHVWRDTIVYLDDNDPILIGRAYGRVSRHLLHEELLRRCVESGVS YLSSKVERIIETTNGQSLIECGTDVVVPCRLATVASGAASGKLLKFEVGGPRVSVQTAYGVEV EVENNPYDPNLMVFMDYRDYAKQKVQPLEAQYPTFLYAMPMSPTRVFFEETCLASKDAMP FDLLKRKLMDRLETMGVHVLKTYEEEWS

Example 20 Construction of a Lutein Producing Strain

The following sequence, optimized for Y. lipolytica codon bias and encoding a putative lycopene epsilon cyclase from Ostreococcus lucimarinus CCE9901, is synthesized de novo:

(SEQ ID NO: 168) TTCTAGAACAAAATGAAGGACGACCGAGAGTGGATCGCCTTCCAGCAGC GAAAGGTGTTCTCTGAGCAGAAGCAGATCAAGGAGTACCTGTCTGCCCT GAACGACCGAGACAAGGTGGACGTGCTGGTGGTGGGCGCCGGCCCCGCC GGCCTGGCCATCGCCGCCGAGACCGCCAAGAAGGGCCTGTCTGTGGGCC TGGTGGCCCCCGACACCCCCTTCGTGAACAACTACGGCGTGTGGCTGGA CGAGTTCAAGGACCTGGGCCTGGAGCACTGTCTGCTGCACAAGTACGAC GACGCCCTGGTGTGGTTCGACGACTCTGACCCCGCCTCTGGCACCGAGC TGGGCCGACCCTACGGCCAGGTGTGTCGACGACGACTGCGAGACCACCT GCTGAAGGAGTGTGCCGCCGCCGGCGTGAAGTACCTGCCCGGCCTGGTG GACTTCGTGCGACACGGCGACGTGGAGAAGAACGAGCTGGCCGAGGCCA ACCGAGGCCAGCAGTTCACCCTGAACTCTCGACTGGTGGTGGCCGGCAC CGGCCACAACCGAGACATGCTGTCTTACGAGGAGGGCGCCCCCCCCGGC TGGCAGACCGCCTACGGCGTGGAGGTGCGAATCCCCAACCACGGCTTCC CCGTGAACAAGGCCGTGTTCATGGACTTCCGACAGTCTGACCCCGAGGC CATGAAGGAGGAGCAGGACGAGGGCGTGTGGCGAGTGCCCTCTTTCCTG TACGTGCTGCCCGTGGACAAGGACGTGGTGTTCGTGGAGGAGACCTGTC TGGTGGCCCGAGTGCAGGTGCCCTTCGACGAGCTGAAGCGACGACTGTA CCGACGAATGAAGCGAATGGGCATGGAGATCGTGGAGGAGGACATCCTG GAGGTGGAGGCCTCTTGGATCCCCCTGGGCGGCACCCCCCCCGTGGCCC CCCAGCGAACCATCGCCTACGGCGCCGCCGCCGGCATGGTGCACCCCGC CTCTGGCTACTCTGTGGTGAACTCTATCTCTAAGGCCCCCCGAGTGGCC ACCGCCATGGCCGAGGGCCTGAAGGAGGGCGGCGAGATCGAGGCCTCTC GACGAGCCTGGGAGATCCTGTGGGGCGCCGAGCCCCGACGACAGATCGG CTTCTACCAGTTCGGCATGGAGCTGCTGATGTCTCTGCGAATCGAGCAG ATGCGAAACTTCTTCTCTACCTTCTTCGCCCTGCCCACCAACCTGTCTC GAGGCTTCCTGGGCAACCGACTGTCTTCTTCTGAGCTGATCATGTTCGC CCTGACCACCTTCGCCATCGGCAACAACGAGCTGCGAGGCCTGCTGCTG GCCCACCTGGTGTCTTAAACGCGT

This fragment, liberated with XbaI and MluI, is cloned into NheI- and MluI-cleaved pMB5082 to produce pEpCyOs1.

A second putative lycopene epsilon cyclase from Ostreococcus lucimarinus CCE9901 is similarly codon-optimized and synthesized de novo:

(SEQ ID NO: 169) TTCTAGAACAAAATGCGAGCCCGACGAGCCCCCGCCGCCCGAGTGACCC GAGCCATCCGAGCCCGAGGCGACGCCGGCACCCGAGCCCGAGACGTGGC CCCCGGCGCCACCCGACGAGGCGCCTCTGCCACCCCCCGAGCCACCCGA CGACCCTCTGCCCGAGAGACCCGACCCGAGCTGTACGGCCTGGACGCCT CTTGGGACCCCCTGACCTCTGGCGACCGACGAGAGTCTGAGGAGTCTCG AACCCCCCTGCCCGAGACCCTGCCCAACGTGCGATGGGGCACCTCTGCC TCTGAGGCCTACGACCTGGTGATCGTGGGCTGTGGCCCCGCCGGCCTGA CCGCCGCCGACGAGGCCTCTAAGCGAGGCCTGCGAGTGGCCCTGATGGA CCCCTCTCCCCTGGCCCCCTGGATGAACAACTACGGCGTGTGGTGTGAC GAGTTCAAGTCTCTGGGCTTCGACGACTGTTACCGAGCCGTGTGGAACA AGGCCCGAGTGATCATCGACGACGGCGACGCCGACGGCAAGATGCTGGA CCGAGCCTACGCCCAGGTGGACCGAAAGAAGCTGAAGCAGAAGCTGATC GCCCGATCTGTGACCCAGGGCGTGGAGTTCGGCATCGCCGCCGTGGACT CTTGTGACAACTCTGACCCCAACCACTCTGTGGTGACCCTGTCTGACGG CCGAAAGGTGTACGCCAAGATGGTGCTGGACGCCACCGGCCACTCTCGA AAGCTGGTGGACTTCGACCGAGACTTCACCCCCGGCTACCAGGCCGCCT TCGGCATCGTGTGTACCGTGGAGAAGCACGACTTCCCCCTGGACACCAT GCTGTTCATGGACTGGCGAGACGAGCACCTGTCTCCCGAGTTCAAGCGA GCCAACGACCGACTGCCCACCTTCCTGTACGCCATGCCCTTCTCTGAGA CCGAGGTGTTCCTGGAGGAGACCTCTCTGGTGGCCCGACCCGGCCTGGA GTTCGACGACCTGAAGCTGAAGCTGAAGGAGCGACTGGACTACCTGGGC GTGAAGGTGACCAAGGTGCACGAGGAGGAGTACTGTCTGATCCCCATGG GCGGCGTGCTGCCCACCTTCCCCCAGCGAACCCTGGGCATCGGCGGCAC CGCCGGCATGGTGCACCCCTCTACCGGCTTCATGGTGGCCAAGACCATG CTGTGTGTGCGAACCCTGGTGGGCACCCTGGACGAGGCCCTGAAGGCCG GCAAGCGAGGCGACATCACCGGCGCCCTGGAGGCCGCCGAGGCCGCCCA GATGAACAACGGCAAGTTCGACGCCGACGCCACCGCCGCCCTGGTGTGG AACTCTATCTGGCCCGAGAACGACCTGCGAATGCGAACCTTCATGTGTT TCGGCATGGAGACCCTGATGCAGCTGGACATCGACGGCACCCGACAGTT CTTCGACACCTTCTTCGACCTGCCCAAGGACGTGTGGGCCGGCTTCCTG TCTTGGCGAATCCAGCCCGTGGGCCTGCTGTCTCTGGGCGTGAACCTGT TCGCCCTGTTCTCTAACTACATGCGAGTGAACTTCGTGAAGTCTGCCCT GCCCTTCATGGGCTCTTTCTTCGCCAACTAAACGCGT

This fragment, liberated with XbaI and MluI, is cloned into NheI- and MluI-cleaved pMB5082 to produce pEpCyOs2.

The following sequence, optimized for Y. lipolytica codon bias and encoding a putative carotene epsilon hydroxylase from Ostreococcus tauri, is synthesized de novo:

(SEQ ID NO: 170) TTCTAGAACAAAATGAAGGACGGCCAGGACGAGGACTCTGACGAGATCT GGGGCGGCCAGCGACACGCCTCTGAGATGAAGACCCCCACCCGACGAAA GGCCCGAACCAAGGCCGAGCGAGAGGCCTCTGCCGCCTCTTACGAGTGG TCTGCCTGGGCCTCTTCTTGTGGCGTGATCTCTGTGGCCATCACCGCCA CCTACTTCCGAATCCTGCGAGAGGTGGACGTGAACGGCGGCGTGTTCCC CGTGGCCGAGCTGGTGGCCCAGCTGGCCCTGATCGCCGGCGCCGCCGTG GGCATGGAGTTCTACGCCCGATACGCCCACAAGCACCTGTGGCACGGCT CTTGGTGGACCATGTCTAACAAGTACCGACAGGAGTGGAACCGACCCAT CTGGCTGCTGCACGAGTCTCACCACCTGCCCCGAGAGGGCGCCTTCGAG GCCAACGACGTGTTCGCCCTGATGAACGGCGTGCCCGCCTTCGCCCTGT GTGCCTTCGGCTTCTTCACCCCCGGCGTGTTCGGCGGCCTGTGTTTCGG CGCCGGCCTGGGCATCACCCTGTTCGGCATCGCCTACATGTACGTGCAC GACGGCCTGGTGCACAAGCGATTCCCCACCGGCCCCCTGGGCAAGCTGC CCGTGATGCGACGAATCGCCGCCGGCCACACCATCCACCACACCGAGGC CTTCGAGGGCGTGCCCTGGGGCCTGTTCCTGGGCATCCAGGAGCTGGCC GCCGTGCCCGGCGGCCTGGAGGAGCTGGAGAAGGTGGTGATCGCCGCCG AGCGAAAGGAGAAGCGAGACGAGCTGGAGCTGGCCCGACGAGCCTCTGT GGGCCTGGTGACCGAGGGCGCCCACATCCCCTCTATGAAGGAGGCCCCC CAGTGTAAGCTGCCCGAGGACCCCTAAACGCGT

This fragment, liberated with XbaI and MluI, is cloned into NheI- and MluI-cleaved pMB5082 to produce pEpHyOs1.

The 1.9 kb KpnI-SacI TEF1p-crtZ fragment from pMB4837 (example 1O) is cloned into KpnI- and SacI-cleaved pMB5082 to create pCrtZ-Ub.

A strain expressing carRP, carB, GGS, and HMG1_(trunc) and auxotrophic for ura3 (MF946; example 2F) is transformed successively, in any order, with the URA3 plasmids pEpCyOs1 (or pEpCyOs2), pEpHyOs1, and pCrtZ-Ub, with the recycling of the ura3 marker between each step, as described in Example 15. Such a strain is expected to produce >1 mg/gDCW lutein. This strain may be further modified by transformation with pMB4789 (erg9[F317I]-3′UTR::URA3), as described in example 2H.

The following tables are referenced throughout the description. Each reference and information designated by each of the Genbank Accession and GI numbers are hereby incorporated by reference in their entirety. The order of genes, polypeptides and sequences presented in the tables is not indicative of their relative importance and/or suitability to any of the embodiments disclosed herein.

TABLE 1 Examples of acetyl-CoA carboxylase polypeptides. ACCESSION GI PROTEIN DESCRIPTION XP_410263 49097606 hypothetical protein AN6126.2 [Aspergillus nidulans FGSC A4] XP_329580 32418204 hypothetical protein [Neurospora crassa] XP_386756 46124405 hypothetical protein FG06580.1 [Gibberella zeae PH-1] XP_367702 39972623 hypothetical protein MG07613.4 [Magnaporthe grisea 70-15] XP_501721 50548503 hypothetical protein [Yarrowia lipolytica] EAK99708 46440402 hypothetical protein CaO19.7466 [Candida albicans SC5314] XP_457211 50413128 unnamed protein product [Debaryomyces hansenii] NP_982612 45184894 AAR071Wp [Eremothecium gossypii] XP_449236 50293649 unnamed protein product [Candida glabrata] NP_593271 19114183 acetyl-coa carboxylase [Schizosaccharomyces pombe] NP_014413 6324343 Acclp [Saccharomyces cerevisiae] XP_455355 50310667 unnamed protein product [Kluyveromyces lactis] T42531 11272737 acetyl-CoA carboxylase (EC 6.4.1.2)—fission yeast (Schizosaccharomyces pombe) AAA20073 171504 acetyl-CoA carboxylase EAL20176 50257469 hypothetical protein CNBF2520 [Cryptococcus neoformans var. neoformans B-3501A] XP_571316 58268320 acetyl-CoA carboxylase, putative [Cryptococcus neoformans var. neoformans JEC21] XP_402244 49076566 hypothetical protein UM04629.1 [Ustilago maydis 521] S60200 2133343 acetyl-CoA carboxylase (EC 6.4.1.2)—smut fungus (Ustilago maydis) BAA24410 2804173 acetyl-coenzyme A carboxylase like carboxylase [Saccharomyces cerevisiae] P32874 1708192 HFA1 protein S55089 7438088 probable acetyl-CoA carboxylase (EC 6.4.1.2) HFA1—yeast (Saccharomyces cerevisiae) NP_990836 45382859 acetyl-Coenzyme A carboxylase alpha [Gallus gallus] CAE01471 32526576 Acetyl-CoA carboxylase 2 [Homo sapiens] AAR37018 40019048 acetyl-CoA carboxylase 2 [Homo sapiens] NP_001 . . . 57164283 acetyl-CoA carboxylase [Ovis aries] NP_776649 27806341 acetyl-coenzyme A carboxylase alpha [acetyl-coA carboxylase] [Bos taurus] CAI25271 56205878 acetyl-Coenzyme A carboxylase [Mus musculus] XP_109883 51828611 PREDICTED: acetyl-Coenzyme A carboxylase [Mus musculus] NP_942134 38679971 acetyl-Coenzyme A carboxylase alpha isoform 3 [Homo sapiens] NP_942131 38679960 acetyl-Coenzyme A carboxylase alpha isoform 1 [Homo sapiens] NP_942135 38679974 acetyl-Coenzyme A carboxylase alpha isoform 4 [Homo sapiens] NP_942136 38679977 acetyl-Coenzyme A carboxylase alpha isoform 2 [Homo sapiens] AAP94122 33112885 acetyl-CoA carboxylase 1 [Homo sapiens] NP_071529 11559962 acetyl-coenzyme A carboxylase alpha [Rattus norvegicus] 2006242A 740964 Ac-CoA carboxylase AAS13685 42405896 acetyl-CoA carboxylase 1 [Mus musculus] NP_598665 48976025 acetyl-Coenzyme A carboxylase beta [Mus musculus] Q13085 2493311 Acetyl-CoA carboxylase 1 (ACC-alpha) [Includes: Biotin carboxylase] XP_548250 57091783 PREDICTED: similar to acetyl-CoA carboxylase [Canis familiaris] XP_314071 58385597 ENSANGP00000015662 [Anopheles gambiae str. PEST] CAG08536 47226520 unnamed protein product [Tetraodon nigroviridis] NP_724636 24586460 CG11198-PB, isoform B [Drosophila melanogaster] NP_610342 24586458 CG11198-PA, isoform A [Drosophila melanogaster] NP_001084 4501855 acetyl-Coenzyme A carboxylase beta [Homo sapiens] NP_446374 16758804 acetyl-Coenzyme A carboxylase beta [Rattus norvegicus] EAL63219 60465120 acetyl-CoA carboxylase [Dictyostelium discoideum] NP_921034 37533464 putative acetyl-CoA carboxylase [Oryza sativa (japonica cultivar- group)] T07084 7438099 acetyl-CoA carboxylase (EC 6.4.1.2) A—soybean AAP78896 32264940 acetyl-coenzyme A carboxylase ACC1A [Zea mays] AAO62903 29123370 acetyl-coenzyme A carboxylase [Setaria italica] BAA07012 1100253 acetyl-CoA carboxylase [Arabidopsis thaliana] AAL02056 15558947 acetyl-coenzyme A carboxylase [Setaria italica] AAG40563 11869927 acetyl-CoA carboxylase 1 [Arabidopsis thaliana] D86483 25293894 protein F5J5.19 [imported]—Arabidopsis thaliana T07920 7438090 probable acetyl-CoA carboxylase (EC 6.4.1.2)—rape A57710 2130099 acetyl-CoA carboxylase (EC 6.4.1.2)—wheat AAO62902 29123376 acetyl-coenzyme A carboxylase [Setaria italica] 2208491A 1588584 Ac-CoA carboxylase T09538 7438102 acetyl-CoA carboxylase (EC 6.4.1.2)—alfalfa CAC19875 12057067 acetyl-CoA carboxylase [Brassica napus] AAP78897 32264942 acetyl-coenzyme A carboxylase ACC1B [Zea mays] T02235 7438095 acetyl-CoA carboxylase (EC 6.4.1.2)—maize AAG40564 11869928 acetyl-CoA carboxylase 2 [Arabidopsis thaliana] E86483 25293893 probable acetyl-CoA carboxylase, 9984-22276 [imported]— Arabidopsis thaliana CAC84161 20975574 acetyl-coenzyme A carboxylase [Alopecurus myosuroides] T07081 7438097 acetyl-CoA carboxylase (EC 6.4.1.2) B—soybean (fragment) CAC19876 12057069 acetyl-CoA carboxylase [Brassica napus]

TABLE 2 Examples of pyruvate decarboxylase polypeptides. ACCESSION GI PROTEIN DESCRIPTION 1QPBB 7245977 Chain B, Pyruvate Decarboyxlase From Yeast (Form B) Complexed With Pyruvamide CAA54522 871533 pyruvate decarboxylase [Saccharomyces cerevisiae] 1PYDB 515237 Chain B, Pyruvate Decarboxylase (Pdc) (E.C.4.1.1.1) CAA28380 4109 pyruvate decarboxylase [Saccharomyces cerevisiae] 1PVDB 1127233 Chain B, Pyruvate Decarboxylase (Pdc) (E.C.4.1.1.1) CAA33709 4114 unnamed protein product [Saccharomyces cerevisiae] AAN77243 25992752 pyruvate decarboxylase [Candida glabrata] NP_013235 6323163 Minor isoform of pyruvate decarboxylase, key enzyme in alcoholic fermentation [Saccharomyces cerevisiae] Q6FJA3 57012668 Pyruvate decarboxylase S36363 486942 pyruvate decarboxylase (EC 4.1.1.1)—yeast (Kluyveromyces marxianus) Q12629 52788279 Pyruvate decarboxylase AAP75898 37359468 pyruvate decarboxylase [Saccharomyces kluyveri] S70684 2131152 pyruvate decarboxylase (EC 4.1.1.1)—yeast (Kluyveromyces marxianus var. lactis) NP_011601 6321524 Minor isoform of pyruvate decarboxylase, key enzyme in alcoholic fermentation [Saccharomyces cerevisiae] AAQ73618 34500072 pyruvate decarboxylase [Saccharomyces kluyveri] NP_983270 45185554 ACL134Cp [Eremothecium gossypii] AAF78895 8745337 putative pyruvate decarboxylase [Saccharomyces kluyveri] CAB65554 6689662 putative pyruvate decarboxylase [Zygosaccharomyces bisporus] AAP75899 37359470 pyruvate decarboxylase [Saccharomyces kluyveri] NP_982469 45184751 AAL073Wp [Eremothecium gossypii] CAA97091 1945321 PDC6 [Saccharomyces cerevisiae] S50700 1086157 pyruvate decarboxylase (EC 4.1.1.1)—yeast (Hanseniaspora uvarum) XP_446491 50288125 unnamed protein product [Candida glabrata] XP_462338 50427451 unnamed protein product [Debaryomyces hansenii] AAC03164 17066784 pyruvate decarboxylase 1 [Pichia stipitis] EAK96569 46437219 hypothetical protein CaO19.10395 [Candida albicans SC5314] XP_457131 50412425 unnamed protein product [Debaryomyces hansenii] AAC03165 2734883 pyruvate decarboxylase 2 [Pichia stipitis] XP_459224 50421349 unnamed protein product [Debaryomyces hansenii] CAH56494 52673248 pyruvate decarboxylase [Pichia anomala] XP_502647 50550349 hypothetical protein [Yarrowia lipolytica] NP_010203 6320123 Probable decarboxylase, [Saccharomyces cerevisiae] BAA04886 1786148 THI3 regulatory protein [Saccharomyces cerevisiae] XP_449074 50293325 unnamed protein product [Candida glabrata] EAL04098 46444826 hypothetical protein CaO19.12078 [Candida albicans SC5314] CAD60727 27803024 unnamed protein product [Podospora anserina] T38759 25777585 probable pyruvate decarboxylase (EC 4.1.1.1)—fission yeast (Schizosaccharomyces pombe) XP_331173 32421459 hypothetical protein [Neurospora crassa] NP_594083 19114995 pyruvate decarboxylase [Schizosaccharomyces pombe] XP_401609 49075036 hypothetical protein UM03994.1 [Ustilago maydis 521] XP_390010 46136637 hypothetical protein FG09834.1 [Gibberella zeae PH-1] XP_409025 49095128 DCPY_EMENI Pyruvate decarboxylase [Aspergillus nidulans FGSC A4] NP_984350 45188127 ADR254Wp [Eremothecium gossypii] AAD16178 4323053 pyruvate decarboxylase [Aspergillus oryzae] P87208 2501326 Pyruvate decarboxylase EAL18331 50255598 hypothetical protein CNBJ2540 [Cryptococcus neoformans var. neoformans B-3501A] XP_567475 58260130 pyruvate decarboxylase, putative [Cryptococcus neoformans var. neoformans JEC21] AAM73540 21666011 pyruvate decarboxylase PdcB [Rhizopus oryzae] AAM73539 21666009 pyruvate decarboxylase PdcA [Rhizopus oryzae] XP_502508 50550071 hypothetical protein [Yarrowia lipolytica] CAA93158 1177659 SPAC3H8.01 [Schizosaccharomyces pombe] XP_412533 49123327 hypothetical protein AN8396.2 [Aspergillus nidulans FGSC A4] P51844 1706333 Pyruvate decarboxylase XP_455842 50311631 unnamed protein product [Kluyveromyces lactis] CAA61155 3688422 pyruvate decarboxylase [Kluyveromyces lactis] XP_444902 50284947 unnamed protein product [Candida glabrata] CAA47319 4118 pyruvate decarboxylase [Saccharomyces cerevisiae]

TABLE 3 Examples of isocitrate dehydrogenase polypeptides. ACCESSION GI PROTEIN DESCRIPTION O13285 3023996 Isocitrate dehydrogenase [NADP], mitochondrial precursor (IDH) (NADP+-specific ICDH) (IDP) (CtIDP1) EAK91676 46432179 hypothetical protein CaO19.5211 [Candida albicans SC5314] O13285 3023996 Isocitrate dehydrogenase [NADP], mitochondrial precursor (IDH) (NADP+-specific ICDH) (IDP) (CtIDP1) EAK94305 46434909 hypothetical protein CaO19.3733 [Candida albicans SC5314] XP_451683 50303483 unnamed protein product [Kluyveromyces lactis] XP_459772 50422415 unnamed protein product [Debaryomyces hansenii] O13294 27805482 Isocitrate dehydrogenase [NADP] peroxisomal (Oxalosuccinate decarboxylase) (IDH) (PS-NADP-IDH) (CtIDP2) XP_460289 50423413 unnamed protein product [Debaryomyces hansenii] XP_390523 46137663 hypothetical protein FG10347.1 [Gibberella zeae PH-1] XP_367343 39971905 hypothetical protein MG07268.4 [Magnaporthe grisea 70-15] XP_323176 32405126 hypothetical protein [Neurospora crassa] XP_445447 50286037 unnamed protein product [Candida glabrata] AAK76730 15027826 mitochondrial NADP-dependent isocitrate dehydrogenase [Aspergillu snidulans] NP_010217 6320137 Idp1p [Saccharomyces cerevisiae] NP_984921 45190667 AER061Cp [Eremothecium gossypii] AAK76731 15027827 peroxisomal NADP-dependent isocitrate dehydrogenase [Aspergillus nidulans] P79089 3023999 Isocitrate dehydrogenase [NADP], mitochondrial precursor (IDH) (NADP+-specific ICDH) (IDP) NP_013275 6323203 Cytosolic NADP-specific isocitrate dehydrogenase [Saccharomyces cerevisiae] XP_407136 49091350 IDHP_ASPNG ISOCITRATE DEHYDROGENASE [NADP], MITOCHONDRIAL PRECURSOR (IDP) [Aspergillus nidulans FGSC A4] NP_982520 45184802 AAL022Wp [Eremothecium gossypii] XP_446953 50289047 unnamed protein product [Candida glabrata] XP_445184 50285511 unnamed protein product [Candida glabrata] XP_455638 50311227 unnamed protein product [Kluyveromyces lactis] AAA64516 736722 isocitrate dehydrogenase NP_970434 42525054 isocitrate dehydrogenase (NADP) [Bdellovibrio bacteriovorus HD100] AAT93173 51013759 YNL009W [Saccharomyces cerevisiae] XP_569233 58264154 isocitrate dehydrogenase (NADP+), putative [Cryptococcus neoformans var. neoformans JEC21] XP_569234 58264156 isocitrate dehydrogenase (NADP+), putative [Cryptococcus neoformans var. neoformans JEC21] XP_403726 49080406 hypothetical protein UM06111.1 [Ustilago maydis 521] XP_503571 50552322 XP_458151 50428131 unnamed protein product [Debaryomyces hansenii] O13302 13124301 Isocitrate dehydrogenase [NAD] subunit 1, mitochondrial precursor (NAD+-specific ICDH) XP_409927 49096934 IDH1_AJECA Isocitrate dehydrogenase [NAD] subunit 1, mitochondrial precursor [Aspergillus nidulans FGSC A4] XP_385909 46122711 IDH1_AJECA Isocitrate dehydrogenase [NAD] subunit 1, mitochondrial precursor [Gibberella zeae PH-1] XP_365293 39967489 hypothetical protein MG01995.4 [Magnaporthe grisea 70-15] NP_983873 45187650 ADL223Wp [Eremothecium gossypii] XP_455266 50310493 IDH1_KLULA [Kluyveromyces lactis] NP_594397 19115309 putative isocitrate dehydrogenase (EC 1.1.1.41) [Schizosaccharomyces pombe] XP_324955 32408949 hypothetical protein [Neurospora crassa] CAE81942 38636405 probable isocitrate dehydrogenase [NAD] subunit 1, mitochondrial precursor [Neurospora crassa] NP_014361 6324291 Idh1p [Saccharomyces cerevisiae] XP_446479 50288101 unnamed protein product [Candida glabrata] XP_567378 58259936 isocitrate dehydrogenase (NAD+), putative [Cryptococcus neoformans var. neoformans JEC21] XP_398944 49069310 hypothetical protein UM01329.1 [Ustilago maydis 521] XP_502479 50550013 hypothetical protein [Yarrowia lipolytica] EAK96238 46436883 hypothetical protein CaO19.13213 [Candida albicans SC5314] EAK96305 46436951 hypothetical protein CaO19.5791 [Candida albicans SC5314] XP_461797 50426401 unnamed protein product [Debaryomyces hansenii] XP_328403 32415850 hypothetical protein [Neurospora crassa] CAF31997 42820684 isocitrate dehydrogenase, putative [Aspergillus fumigatus] XP_389756 46136129 hypothetical protein FG09580.1 [Gibberella zeae PH-1] XP_363786 39952139 hypothetical protein MG01712.4 [Magnaporthe grisea 70-15] AAL73035 18463935 isocitrate dehydrogenase [Coccidioides immitis] XP_405140 49086142 hypothetical protein AN1003.2 [Aspergillus nidulans FGSC A4] NP_595203 19111995 probable isocitrate dehydrogenase [nad] subunit 2, mitochondrial precursor [Schizosaccharomyces pombe] NP_014779 6324709 Idh2p [Saccharomyces cerevisiae] XP_447564 50290265 unnamed protein product [Candida glabrata] NP_985684 45198655 AFR137Cp [Eremothecium gossypii] XP_566837 58258849 isocitrate dehydrogenase, putative [Cryptococcus neoformans var. neoformans JEC21] XP_454086 50308171 IDH2_KLULA [Kluyveromyces lactis] XP_398943 49069308 hypothetical protein UM01328.1 [Ustilago maydis 521]

TABLE 4 Examples of ATP-citrate lyase polypeptides. ACCESSION GI PROTEIN DESCRIPTION XP_327071 32413182 hypothetical protein [Neurospora crassa] O93988 30912679 ATP-citrate synthase subunit 1 (ATP-citrate (pro-S-)-lyase 1) (Citrate cleavage enzyme subunit 1) XP_370222 39977669 hypothetical protein MG06719.4 [Magnaporthe grisea 70-15] XP_406573 49090008 ACL1_NEUCR Probable ATP-citrate synthase subunit 1 [Aspergillus nidulans FGSC A4] XP_504787 50554757 hypothetical protein [Yarrowia lipolytica] Q9P7W3 30912748 Probable ATP-citrate synthase subunit 1 (ATP-citrate (pro-S-)-lyase 1) (Citrate cleavage enzyme subunit 1) XP_398620 49068662 hypothetical protein UM01005.1 [Ustilago maydis 521] NP_596202 19112994 probable ATP citrate lyase [Schizosaccharomyces pombe] XP_567460 58260100 conserved hypothetical protein [Cryptococcus neoformans var. neoformans JEC21] NP_001008 56118260 acly-prov protein [Xenopus tropicalis] XP_418154 50760837 PREDICTED: similar to ATP citrate lyase [Gallus gallus] AAH84253 54038148 LOC495086 protein [Xenopus laevis] NP_942127 38569423 ATP citrate lyase isoform 2 [Homo sapiens] NP_001087 38569421 ATP citrate lyase isoform 1 [Homo sapiens] P53396 20141248 ATP-citrate synthase (ATP-citrate (pro-S-)-lyase) (Citrate cleavage enzyme) AAL34316 17028103 ATP-citrate lyase [Rattus norvegicus] NP_001002 50540366 zgc:92008 [Danio rerio] AAH84776 54311201 LOC495316 protein [Xenopus laevis] S21173 105392 ATP citrate (pro-S)-lyase—human AAT94429 51092031 RE70805p [Drosophila melanogaster] AAD34754 28372804 LD21334p [Drosophila melanogaster] AAH21502 18204829 Acly protein [Mus musculus] XP_319323 58392375 ENSANGP00000012364 [Anopheles gambiae str. PEST] NP_725514 24653990 CG8322-PB, isoform B [Drosophila melanogaster] EAL26601 54637198 GA20986-PA [Drosohilia pseudoobscura] CAE56725 39579419 Hypothetical protein CBG24512 [Caenorhabditis briggsae] CAE64663 39593194 Hypothetical protein CBG09435 [Caenorhabditis briggsae] XP_511495 55645405 PREDICTED: similar to ATP citrate lyase isoform 1 [Pan troglodytes] CAF95829 47210997 unnamed protein product [Tetraodon nigroviridis] AAO22565 27754223 putative ATP citrate lyase [Arabidopsis thaliana] AAL33788 17065616 putative ATP citrate lyase [Arabidopsis thaliana] CAB46077 5304837 ATP citrate lyase [Cyanophora paradoxa] CAF96044 47204726 unnamed protein product [Tetraodon nigroviridis] AAK13318 13160653 ATP: citrate lyase [Capsicum annuum] AAQ75159 34558815 citrate lyase subunit 2 [Alvinella pompejana epibiont 7G3] AAQ75128 34558783 citrate lyase subunit 2 [Alvinella pompejana epibiont 6C6] XP_537640 57091075 PREDICTED: similar to ATP citrate lyase isoform 2 [Canis familiaris] XP_327069 32413178 hypothetical protein ( (AJ243817) ATP citrate lyase, subunit 2 [Sordaria macrospora]) [Neurospora crassa] CAB76164 7160184 ATP citrate lyase, subunit 2 [Sordaria macrospora] XP_370223 39977671 hypothetical protein MG06720.4 [Magnaporthe grisea 70-15] XP_386215 46123323 hypothetical protein FG06039.1 [Gibberella zeae PH-1] CAA10666 7159697 ATP-citrat-lyase [Gibberella pulicaris] XP_406572 49090004 conserved hypothetical protein [Aspergillus nidulans FGSC A4] XP_503231 50551515 hypothetical protein [Yarrowia lipolytica] NP_593246 19114158 putative ATP-citrate (pro-S-) lyase (EC 4.1.3.8) [Schizosaccharomyces pombe] XP_398620 49068662 hypothetical protein UM01005.1 [Ustilago maydis 521] XP_567460 58260100 conserved hypothetical protein [Cryptococcus neoformans var. neoformans JEC21] AAT94429 51092031 RE70805p [Drosophila melanogaster] NP_725514 24653990 CG8322-PB, isoform B [Drosophila melanogaster] AAD34754 28372804 LD21334p [Drosophila melanogaster] EAL26601 54637198 GA20986-PA [Drosophila pseudoobscura] XP_319323 58392375 ENSANGP00000012364 [Anopheles gambiae str. PEST] AAH84776 54311201 LOC495316 protein [Xenopus laevis] BAB00624 9229902 ATP citrate-lyase [Ciona intestinalis] NP_001008 56118260 acly-prov protein [Xenopus tropicalis] AAH84253 54038148 LOC495086 protein [Xenopus laevis] AAH56378 38614162 ATP citrate lyase [Mus musculus] NP_001087 38569421 ATP citrate lyase isoform 1 [Homo sapiens] NP_942127 38569423 ATP citrate lyase isoform 2 [Homo sapiens] P53396 20141248 ATP-citrate synthase (ATP-citrate (pro-S-)-lyase) (Citrate cleavage enzyme) XP_511495 55645405 PREDICTED: similar to ATP citrate lyase isoform 1 [Pan troglodytes] NP_058683 8392839 ATP citrate lyase [Rattus norvegicus] NP_001002 50540366 zgc: 92008 [Danio rerio] S21173 105392 ATP citrate (pro-S)-lyase—human NP_508280 17551266 atp citrate lyase (XC101) [Caenorhabditis elegans] CAE64663 39593194 Hypothetical protein CBG09435 [Caenorhabditis briggsae] CAE56725 39579419 Hypothetical protein CBG24512 [Caenorhabditis briggsae] NP_506267 17557344 ATP citrate lyase (120.6 kD) (5N599) [Caenorhabditis elegans] XP_537640 57091075 PREDICTED: similar to ATP citrate lyase isoform 2 [Canis familiaris] CAF96059 47204551 unnamed protein product [Tetraodon nigroviridis] F96633 25404292 hypothetical protein F8A5.32 [imported]—Arabidopsis thaliana AAM91141 22136126 similar to ATP-citrate-lyase [Arabidopsis thaliana] NP_849634 30681854 expressed protein [Arabidopsis thaliana] AAO23582 27764922 At1g09430/F19J9_9 [Arabidopsis thaliana] AAM65078 21593129 ATP citrate-lyase, putative [Arabidopsis thaliana] CAC86996 15919089 ATP citrate lyase b-subunit [Lupinus albus] AAQ75158 34558814 citrate lyase subunit 1 [Alvinella pompejana epibiont 7G3] AAQ75127 34558782 citrate lyase subunit 1 [Alvinella pompejana epibiont 6C6]

TABLE 5 Examples of malic enzyme polypeptides. ACCESSION GI PROTEIN DESCRIPTION NP_012896 6322823 Mae1p [Saccharomyces cerevisiae] XP_448858 50292851 unnamed protein product [Candida glabrata] XP_454793 50309563 unnamed protein product [Kluyveromyces lactis] NP_986598 45201028 AGL068Wp [Eremothecium gossypii] XP_460887 50424595 unnamed protein product [Debaryomyces hansenii] EAK97738 46438407 hypothetical protein CaO19.3419 [Candida albicans SC5314] XP_504112 50553402 hypothetical protein [Yarrowia lipolytica] XP_330094 32419237 hypothetical protein [Neurospora crassa] XP_380981 46107844 hypothetical protein FG00805.1 [Gibberella zeae PH-1] XP_411070 49102552 hypothetical protein AN6933.2 [Aspergillus nidulans FGSC A4] XP_362875 39946676 hypothetical protein MG08562.4 [Magnaporthe grisea 70-15] NP_587760 19075260 malate oxidoreductase [nad] [Schizosaccharomyces pombe] NP_978189 42780942 malate oxidoreductase [Bacillus cereus ATCC 10987] YP_035982 49481098 NAD-dependent malic enzyme [Bacillus thuringiensis serovar konkukian str. 97-27] YP_027934 49184682 malate oxidoreductase [Bacillus anthracis str. Sterne] YP_018438 47527089 malate oxidoreductase [Bacillus anthracis str. ‘Ames Ancestor’] ZP_002365 47565532 malate oxidoreductase VC1188 [Bacillus cereus G9241] YP_083209 52143619 NAD-dependent malic enzyme [Bacillus cereus ZK] XP_571672 58269032 malate dehydrogenase, putative [Cryptococcus neoformans var. neoformans JEC21] NP_391586 16080758 hypothetical protein BSU37050 [Bacillus subtilis subsp. subtilis str. 168] YP_092693 52786864 MalS [Bacillus licheniformis ATCC 14580] NP_831516 30019885 NAD-dependent malic enzyme [Bacillus cereus ATCC 14579] YP_093460 52787631 YwkA [Bacillus licheniformis ATCC 14580] YP_081030 52082239 Malic oxidoreductase [Bacillus licheniformis ATCC 14580] NP_822689 29828055 putative malate dehydrogenase [Streptomyces avermitilis MA-4680] O34389 33517449 Probable NAD-dependent malic enzyme 3 (NAD-ME 3) EAL19111 50256386 hypothetical protein CNBH2110 [Cryptococcus neoformans var. neoformans B-3501A] NP_825047 29830413 putative malate dehydrogenase [Streptomyces avermitilis MA-4680] ZP_002340 47096498 NADP-dependent malic enzyme [Listeria monocytogenes str. 1/2a F6854] NP_928837 37525493 malate dehydrogenase (oxaloacetate-decarboxylating) [Photorhabdus luminescens subsp. laumondii TTO1] NP_230833 15641201 malate oxidoreductase [Vibrio cholerae O1 biovar eltor str. N16961] NP_934257 37679648 malic enzyme [Vibrio vulnificus YJ016] NP_761613 27366085 Malic enzyme [Vibrio vulnificus CMCP6] AC1314 25283688 malolactic enzyme (malate dehydrogenase) homolog lmo1915 - Listeria monocytogenes (strain EGD-e) YP_055602 50842375 NAD-dependent malic enzyme [Propionibacterium acnes KPA171202] YP_095310 52841511 malate oxidoreductase [Legionella pneumophila subsp. pneumophila str. Philadelphia 1] ZP_002315 47093832 NADP-dependent malic enzyme [Listeria monocytogenes str. 4b H7858] AC1686 25283689 malolactic enzyme (malate dehydrogenase) homolog lin2029 [imported] - Listeria innocua (strain Clip11262) YP_126594 54294179 malate oxidoreductase [Legionella pneumophila str. Lens] YP_123567 54297198 malate oxidoreductase [Legionella pneumophila str. Paris] EAJ76260 44510091 unknown [environmental sequence] YP_114273 53803890 malate oxidoreductase [Methylococcus capsulatus str. Bath] NP_797637 28898032 malate oxidoreductase [Vibrio parahaemolyticus RIMD 2210633] YP_040250 49483026 putative malolactic enzyme [Staphylococcus aureus subsp. aureus MRSA252] ZP_001276 53693400 COG0281: Malic enzyme [Pseudomonas syringae pv. syringae B728a] YP_044961 50083451 NAD-linked malate dehydrogenase, Rossman fold [Acinetobacter sp. ADP1] YP_128226 54295811 hypothetical protein lpl2901 [Legionella pneumophila str. Lens] NP_719387 24375344 malate oxidoreductase [Shewanella oneidensis MR-1] XP_572853 58271394 nad-dependent malic enzyme, putative [Cryptococcus neoformans var. neoformans JEC21] NP_252161 15598667 probable malic enzyme [Pseudomonas aeruginosa PAO1] ZP_001368 46164263 COG0281: Malic enzyme [Pseudomonas aeruginosa UCBPP-PA14] YP_125345 54298976 hypothetical protein lpp3043 [Legionella pneumophila str. Paris] NP_793695 28871076 malate dehydrogenase [Pseudomonas syringae pv. tomato str. DC3000] YP_096964 52843165 malate dehydrogenase (NAD-linked) [Legionella pneumophila subsp. pneumophila str. Philadelphia 1] EAH92280 44245125 unknown [environmental sequence] YP_154988 56459707 Malic enzyme [Idiomarina loihiensis L2TR] EAI68195 44354928 unknown [environmental sequence] YP_070054 51595863 NAD-dependent malic enzyme [Yersinia pseudotuberculosis IP 32953] YP_133025 54303032 putative malate oxidoreductase [Photobacterium profundum SS9] NP_969623 42524243 NAD-dependent malic enzyme [Bdellovibrio bacteriovorus HD100] NP_856009 31793516 PROBABLE [NAD] DEPENDENT MALATE OXIDOREDUCTASE MEZ (MALIC ENZYME) (NAD-MALIC ENZYME) (MALATE DEHYDROGENASE TATE DECARBOXY ATING)) PYRUVIC-MALIC CARBOXYLASE) (NAD-ME) [Mycobacterium bovis AF2122/97] NP_935035 37680426 malic enzyme [Vibrio vulnificus YJ016] YP_050922 50121755 NAD-dependent malic enzyme [Erwinia carotovora subsp. atroseptica SCRI1043] E70705 7431223 probable malate oxidoreductase - Mycobacterium tuberculosis (strain H37RV) NP_216848 57116971 PROBABLE [NAD] DEPENDENT MALATE OXIDOREDUCTASE MEZ (MALIC ENZYME) (NAD-MALIC ENZYME) (MALATE DEHYDROGENASE TATE DECARBOXY ATING)) PYRUVIC-MALIC CARBOXYLASE) (NAD-ME) [Mycobacterium tuberculosis H37Rv] YP_143786 55980489 NAD-dependent malic enzyme (malate dehydrogenase) [Thermus thermophilus HB8] YP_130202 54309182 putative malate oxidoreductase [Photobacterium profundum SS9] NP_415996 16129438 NAD-linked malate dehydrogenase [Escherichia coli K12] NP_819843 29654151 malate oxidoreductase [Coxiella burnetii RSA 493] NP_753809 26247769 NAD-dependent malic enzyme [Escherichia coli CFT073] NP_707611 56479957 NAD-linked malate dehydrogenase (malic enzyme) [Shigella flexneri 2a str. 301] F85728 25283682 NAD-linked malate dehydrogenase (malic enzyme) - Escherichia coli (strain O157:H7, substrain EDL933) YP_163690 56552851 malic enzyme [Zymomonas mobilis subsp. mobilis ZM4] YP_150562 56413487 NAD-linked malic enzyme [Salmonella enterica subsp. enterica serovar Paratypi A str. ATCC 9150] NP_720610 24378655 malolactic enzyme [Streptococcus mutans UA159] NP_460525 16764910 NAD-linked malate dehydrogenase [Salmonella typhimurium LT2] ZP_003193 48865537 COG0281: Malic enzyme [Oenococcus oeni PSU-1] NP_784797 28377905 malolactic enzyme [Lactobacillus plantarum WCFS1] T13496 7431227 malolactic enzyme (EC 1.1.1.—) - Leuconostoc oenos AAV65766 55793550 malolactic enzyme [Oenococcus oeni] A97096 25283683 malic enzyme [imported] - Clostridium acetobutylicum YP_193951 58337366 malolactic enzyme [Lactobacillus acidophilus NCFM] H97096 25283684 malic enzyme [imported] - Clostridium acetobutylicum ZP_003237 48870993 COG0281: Malic enzyme [Pediococcus pentosaceus ATCC 25745] ZP_001460 41689468 COG0281: Malic enzyme [Psychrobacter sp. 273-4] D86737 25283676 malolactic enzyme [imported] - Lactococcus lactis subsp. lactis (strain IL1403) ZP_002870 48825851 COG0281: Malic enzyme [Enterococcus faecium] ZP_001439 34762975 Malolactic enzyme [Fusobacterium nucleatum subsp. vincentii ATCC 49256] 1922245A 737262 malolactic enzyme YP_169914 56708018 NAD-dependent malic enzyme [Francisella tularensis subsp. tularensis Schu 4] YP_055027 50841800 putative malate oxidoreductase [Propionibacterium acnes KPA171202] ZP_000625 23023297 COG0281: Malic enzyme [Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293] NP_296302 15807565 malate oxidoreductase [Deinococcus radiodurans R1] NP_285599 15807938 malate oxidoreductase [Deinococcus radiodurans R1] YP_132069 54302076 hypothetical malate oxidoreductase [Photobacterium profundum SS9] CAA50716 467569 malolactic enzyme [Lactococcus lactis] ZP_002906 48833596 COG0281: Malic enzyme [Magnetococcus sp. MC-1] ZP_003155 48861632 COG0281: Malic enzyme [Microbulbifer degradans 2-40] NP_773109 27381580 malic enzyme [Bradyrhizobium japonicum USDA 110] AAQ95658 37622953 malic enzyme [Dictyostelium discoideum] CAC19505 56204311 malic enzyme 1, NADP(+)-dependent, cytosolic [Homo sapiens] AAH80660 51873855 Mod1 protein [Mus musculus] P40927 729986 NADP-dependent malic enzyme (NADP-ME) AAT02533 46850200 NADP-dependent malic enzyme 1 [Hydrilla verticillata] BAC37086 26346875 unnamed protein product [Mus musculus] T02763 7431235 probable malate dehydrogenase (oxaloacetate-decarboxylating) (NADP) (EC 1.1.1.40) - maize XP_387367 46125627 hypothetical protein FG07191.1 [Gibberella zeae PH-1] AAC50613 1465733 cytosolic NADP(+)-dependent malic enzyme CAA39421 669118 sbcA8 recE fusion [Escherichia coli] CAA39420 669117 sbcA8 recE fusion [Escherichia coli] NP_032641 6678912 malic enzyme, supernatant [Mus musculus] CAA39419 581228 sbcA8 recE fusion [Escherichia coli] AAB01380 1335389 NADP-dependent malic enzyme JC4160 1085347 malate dehydrogenase (oxaloacetate-decarboxylating) (NADP) (EC 1.1.1.40) - human E96828 25283677 probable malate oxidoreductase, 93001-96525 [imported] - Arabidopsis thaliana BAD87910 57899974 cytosolic NADP malic enzyme [Oryza sativa (japonica cultivar-group)] EAJ77083 44511304 unknown [environmental sequence] P13697 266504 NADP-dependent malic enzyme (NADP-ME) (Malic enzyme 1) NP_036732 7106353 malic enzyme 1 [Rattus norvegicus] YP_065939 51246055 related to NAD-dependent malic enzyme [Desulfotalea psychrophila LSv54] CAC18164 16944467 related to malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+) [Neurospora crassa] XP_322953 32404680 hypothetical protein [Neurospora crassa] AAK91502 18460985 NADP-dependent malic enzyme [Zea mays] AAQ88396 37147841 non-photosynthetic NADP-malic enzyme [Zea mays] NP_001003 57525624 zgc:100941 [Danio rerio] 1GQ2P 21465488 Chain P, Malic Enzyme From Pigeon Liver AAO26053 28195290 malic enzyme [Mucor circinelloides] AAH84250 54038006 Me2 protein [Xenopus laevis] XP_362590 39946106 hypothetical protein MG08173.4 [Magnaporthe grisea 70-15] AAH03287 13096987 Mod1 protein [Mus musculus] Q29558 2497785 NADP-dependent malic enzyme (NADP-ME) (Malic enzyme 1) XP_532217 57094622 PREDICTED: similar to malate dehydrogenase decarboxylase (NADP+) [Canis familiaris] P28227 126734 NADP-dependent malic enzyme (NADP-ME) NP_496968 17537199 malic enzyme nadp-dependent (2O518) [Caenorhabditis elegans] NP_914533 34906372 unnamed protein product [Oryza sativa (japonica cultivar-group)] AAD10504 4096786 NADP-malic enzyme [Zea mays] AAO67523 50897495 mitochondrial malic enzyme 2 [Xenopus laevis] P43279 1170871 NADP-dependent malic enzyme, chloroplast precursor (NADP-ME) AAK83074 15077109 putative cytosolic NADP-malic enzyme [Flaveria pringlei] AAP33011 30575690 NADP-malic enzyme [Zea mays] AAN86690 27357017 malic enzyme [Mastigamoeba balamuthi] P78715 41017288 Malic enzyme, hydrogenosomal precursor (ME) AAP32204 30526303 NADP-dependent malic enzyme [Sorghum bicolor] AAV31249 54287505 NADP malic enzyme [Oryza sativa (japonica cultivar-group)] T06402 7431232 malate dehydrogenase (oxaloacetate-decarboxylating) (NADP) (EC 1.1.1.40) 2, cytosolic - tomato Q99KE1 55583978 NAD-dependent malic enzyme, mitochondrial precursor (NAD-ME) (Malic enzyme 2) XP_399922 49071266 hypothetical protein UM02307.1 [Ustilago maydis 521] P36444 547886 NADP-dependent malic enzyme, chloroplast precursor (NADP-ME) AAO30034 28059162 malate oxidoreductase (malic enzyme) [Arabidopsis thaliana] AAK83073 15077107 putative cytosolic NADP-malic enzyme [Flaveria pringlei] NP_002387 4505145 malic enzyme 2, NAD(+)-dependent, mitochondrial [Homo sapiens] AAA33487 168528 NADP-dependent malic enzyme (EC 1.1.1.40) BAA74735 4239891 NADP-malic enzyme [Aloe arborescens] NP_989634 45383538 malic enzyme 1, NADP(+)-dependent, cytosolic [Gallus gallus] 1GZ3D 31615316 Chain D, The Regulation Of Human Mitochondrial Nad(P)+- Dependent Malic Enzyme By Atp And Fumarate AAW56450 57791240 chloroplast NADP-dependent malic enzyme precursor [Flaveria bidentis] AAT02534 46850202 NADP-dependent malic enzyme 2 [Hydrilla verticillata] S29742 422339 malate dehydrogenase (oxaloacetate-decarboxylating) (NADP) (EC 1.1.1.40) - pig roundworm 1O0SB 34811253 Chain B, Crystal Structure Of Ascaris Suum Malic Enzyme Complexed With Nadh P27443 126732 NAD-dependent malic enzyme, mitochondrial precursor (NAD-ME) T06401 7431231 malate dehydrogenase (oxaloacetate-decarboxylating) (NADP) (EC 1.1.1.40) precursor - tomato AAL16175 16226466 AT5g11670/T22P22_60 [Arabidopsis thaliana] AAF73006 8118507 NADP-dependent malic protein [Ricinus communis] AAK97530 15420975 malic enzyme [Meleagris gallopavo] EAI90348 44385841 unknown [environmental sequence] P51615 1708924 NADP-DEPENDENT MALIC ENZYME (NADP-ME) AAA19575 169327 NADP-dependent malic enzyme S43718 1084300 malate dehydrogenase (oxaloacetate-decarboxylating) (NADP) (EC 1.1.1.40) - common ice plant P34105 1346485 NADP-DEPENDENT MALIC ENZYME (NADP-ME) AAS38597 42733630 similar to Mastigamoeba balamuthi Malic enzyme (EC 1.1.1.38) [Dictyostelium discoideum] BAC54101 27530932 cytosolic NADP-malic enzyme [Lithospermum erythrorhizon] AAT02535 46850204 NADP-dependent malic enzyme 3 [Hydrilla verticillata] CAB66003 6706333 NADP-dependent malate dehydrogenase (decarboxylating) [Apium graveolens] AAH84860 54311418 LOC495390 protein [Xenopus laevis] CAA39422 669119 sbcA8 recE fusion [Escherichia coli] NP_916713 34910732 P0022F10.12 [Oryza sativa (japonica cultivar-group)] CAA56354 510876 NADP dependent malic enzyme [Phaseolus vulgaris] DEFBC 7427668 malate dehydrogenase (oxaloacetate-decarboxylating) (NADP) (EC 1.1.1.40) - kidney bean JC5967 7431234 malate dehydrogenase (oxaloacetate-decarboxylating) (NADP) (EC 1.1.1.40) - aloe NP_197960 15239517 malate oxidoreductase, putative [Arabidopsis thaliana] NP_651959 21356279 CG5889-PA [Drosophila melanogaster] CAB64263 6634090 malate dehydrogenase (NADP-dependent oxaloacetate decarboxylating), malic enzyme [Drosophila melanogaster] BAB20887 54606800 NADP dependent malic enzyme [Oryza sativa (japonica cultivar- group)] EAL27424 54638022 GA19206-PA [Drosophila pseudoobscura] NP_006671 5729920 malic enzyme 3, NADP(+)-dependent, mitochondrial [Homo sapiens] AAB08874 1561774 malate dehydrogenase [Vitis vinifera] 1PJLH 33358128 Chain H, Crystal Structure Of Human M-Nad-Me In Ternary Complex With Nad And Lu3+ 1GZ4D 22218682 Chain D, The Regulation Of Human Mitochondrial Nad(P)+- Dependent Malic Enzyme By Atp And Fumarate 1QR6B 5822327 Chain B, Human Mitochondrial Nad(P)-Dependent Malic Enzyme 1PJ3D 39654475 Chain D, Crystal Structure Of Human Mitochondrial Nad(P)+- Dependent Malic Enzyme P22178 126736 NADP-dependent malic enzyme, chloroplast precursor (NADP-ME) XP_410305 49097690 hypothetical protein AN6168.2 [Aspergillus nidulans FGSC A4] AAH22472 18490280 Malic enzyme 3, NADP(+)-dependent, mitochondrial [Homo sapiens]

TABLE 6 Examples of AMP deaminase polypeptides. ACCESSION GI PROTEIN DESCRIPTION AAA34420 171053 AMP deaminase (EC 3.5.4.6) XP_446684 50288509 unnamed protein product [Candida glabrata] NP_983153 45185436 ABR204Cp [Eremothecium gossypii] XP_453337 50306727 unnamed protein product [Kluyveromyces lactis] EAL02322 46443037 adenosine/AMP deaminase [Candida albicans SC5314] XP_460211 50423261 unnamed protein product [Debaryomyces hansenii] XP_503822 50552824 hypothetical protein [Yarrowia lipolytica] XP_413009 49131023 hypothetical protein AN8872.2 [Aspergillus nidulans FGSC A4] XP_360256 39941438 hypothetical protein MG05630.4 [Magnaporthe grisea 70-15] XP_381547 46108978 hypothetical protein FG01371.1 [Gibberella zeae PH-1] XP_330167 32419447 probable AMP deaminase [MIPS] [Neurospora crassa] CAB97316 16945394 probable AMP deaminase [Neurospora crassa] T50996 11359582 probable AMP deaminase [imported] - Neurospora crassa NP_595153 19111945 amp deaminase [Schizosaccharomyces pombe] EAL22226 50259553 hypothetical protein CNBC3640 [Cryptococcus neoformans var. neoformans B-3501A] XP_402237 49076548 hypothetical protein UM04622.1 [Ustilago maydis 521] CAA62797 995562 AMP deaminase [Schizosaccharomyces pombe] AAF65407 7638159 AMP deaminase [Dictyostelium discoideum] XP_537039 57088163 PREDICTED: similar to AMP deaminase 2 (AMP deaminase isoform L) [Canis familiaris] AAH49119 29145073 Adenosine monophosphate deaminase 2 (isoform L) [Mus musculus] XP_569691 58265070 AMP deaminase, putative [Cryptococcus neoformans var. neoformans JEC21] AAD56303 5922018 AMP deaminase isoform L [Homo sapiens] NP_004028 21264318 adenosine monophosphate deaminase 2 (isoform L) isoform 1 [Homo sapiens] A44313 345738 AMP deaminase (EC 3.5.4.6) isoform L - human CAI19307 56206061 adenosine monophosphate deaminase 2 (isoform L) [Homo sapiens] AAA62126 644509 AMP deaminase isoform L splicing variant CAI19305 56206059 adenosine monophosphate deaminase 2 (isoform L) [Homo sapiens] XP_310497 58424203 ENSANGP00000017310 [Anopheles gambiae str. PEST] CAI19306 56206060 adenosine monophosphate deaminase 2 (isoform L) [Homo sapiens] AAC50308 608499 AMP deaminase CAG06825 47229629 unnamed protein product [Tetraodon nigroviridis] NP_727741 45551453 CG32626-PD, isoform D [Drosophila melanogaster] NP_727739 45551452 CG32626-PA, isoform A [Drosophila melanogaster] NP_727740 24641890 CG32626-PB, isoform B [Drosophila melanogaster] AAN09337 22832227 CG32626-PC, isoform C [Drosophila melanogaster] T01259 7484807 AMP deaminase homolog F16M14.21 - Arabidopsis thaliana XP_506591 51963676 PREDICTED P0034A04.129 gene product [Oryza sativa (japonica cultivar-group)] NP_850294 30687456 AMP deaminase, putative/myoadenylate deaminase, putative [Arabidopsis thaliana] CAG07509 47228777 unnamed protein product [Tetraodon nigroviridis] NP_494974 32564190 Adenosine/AMP deaminase (85.4 kD) (2F499) [Caenorhabditis elegans] T15771 7497030 hypothetical protein C34F11.3 - Caenorhabditis elegans CAE59064 39596837 Hypothetical protein CBG02349 [Caenorhabditis briggsae] NP_494973 32564194 Adenosine/AMP deaminase family member (2F499) [Caenorhabditis elegans] BAA06505 1321635 erythrocyte-type AMP deaminase [Homo sapiens] NP_000471 4502079 adenosine monophosphate deaminase (isoform E) [Homo sapiens] S68147 2134756 AMP deaminase (EC 3.5.4.6), erythrocte, splice form 1c - human AAH56380 38614134 Ampd3 protein [Mus musculus] O08739 2494043 AMP DEAMINASE 3 (AMP DEAMINASE ISOFORM E) (AMP DEAMINASE H-TYPE) (HEART-TYPE AMPD) NP_113732 13928736 adenosine monophosphate deaminase 3 [Rattus norvegicus] O09178 2494044 AMP deaminase 3 (AMP deaminase isoform E) XP_420973 50747746 PREDICTED: similar to AMP deaminase 3 (Erythrocyte AMP deaminase) [Gallus gallus] NP_956142 41054127 AMP deaminase 3 [Danio rerio] CAG01709 47222742 unnamed protein product [Tetraodon nigroviridis] NP_957187 41053780 hypothetical protein MGC77905 [Danio rerio] XP_392957 48104570 similar to ENSANGP00000017310 [Apis mellifera] AAH07183 13938134 Ampd3 protein [Mus musculus] CAG05605 47220579 unnamed protein product [Tetraodon nigroviridis] NP_620231 20302047 adenosine monophosphate deaminase 1 (isoform M) [Rattus norvegicus] XP_540247 57098851 PREDICTED: similar to AMP deaminase 1 (AMP deaminase isoform M) [Canis familiaris] CAF99638 47230445 unnamed protein product [Tetraodon nigroviridis] XP_513671 55587796 PREDICTED: adenosine monophosphate deaminase 1 (isoform M) [Pan troglodytes] CAI18828 56203368 adenosine monophosphate deaminase 1 (isoform M) [Homo sapiens] CAI18829 56203369 OTTHUMP00000059283 [Homo sapiens] CAI18830 56203370 adenosine monophosphate deaminase 1 (isoform M) [Homo sapiens] EAA19931 23484684 AMP deaminase homolog [Plasmodium yoelii yoelii] CAH99706 56500932 AMP deaminase, putative [Plasmodium berghei] XP_131103 38076931 similar to AMP deaminase 1 (Myoadenylate deaminase) (AMP deaminase isoform M) [Mus musculus] CAH77387 56523366 AMP deaminase, putative [Plasmodium chabaudi]

TABLE 7 Examples of acetoacetyl-CoA thiolase polypeptides. ACCESSION GI PROTEIN DESCRIPTION P10551 135758 Acetyl-CoA acetyltransferase (Acetoacetyl-CoA thiolase) Q04677 418002 Acetyl-CoA acetyltransferase IB (Peroxisomal acetoacetyl-CoA thiolase) (Thiolase IB) Q12598 34925109 Acetyl-CoA acetyltransferase IA (Peroxisomal acetoacetyl-CoA thiolase) (Thiolase IA) T10247 7433657 acetyl-CoA C-acetyltransferase (EC 2.3.1.9), cytosolic - radish T42741 11257345 probable acetyl-CoA C-acetyltransferase (EC 2.3.1.9) - fission yeast (Schizosaccharomyces pombe) AAL18924 16417944 acetyl Co-A acetyltransferase [Hevea brasiliensis] AAM67058 21618008 acetoacyl-CoA-thiolase [Arabidopsis thaliana] AAO51605 28829030 similar to acetyl-coa acetyltransferase (EC 2.3.1.9) [Schizosaccharomyces pombe] AAU95618 53854350 cytosolic acetoacetyl-coenzyme A thiolase [Nicotiana tabacum] AAU95619 53854352 peroxisomal acetoacetyl-coenzyme A thiolase [Nicotiana tabacum] BAA97003 8777413 acetyl-CoA C-acetyltransferase [Arabidopsis thaliana] CAE76429 38567134 probable acetoacetyl-CoA thiolase [Neurospora crassa] EAK90852 46431255 hypothetical protein CaO19.1591 [Candida albicans SC5314] EAL32264 54643520 GA10651-PA [Drosophila pseudoobscura] NP_015297 6325229 Acetyl-CoA C-acetyltransferase (acetoacetyl-CoA thiolase) [Saccharomyces cerevisiae] NP_568694 30695411 acetyl-CoA C-acyltransferase, putative/3-ketoacyl-CoA thiolase, putative [Arabidopsis thaliana] NP_572414 24640423 CG10932-PA [Drosophila melanogaster] NP_596686 19113478 acetyl-coa acetyltransferase (EC 2.3.1.9) [Schizosaccharomyces pombe] NP_851154 30695409 acetyl-CoA C-acyltransferase, putative/3-ketoacyl-CoA thiolase, putative [Arabidopsis thaliana] NP_908411 34894172 putative acetoacetyl-coenzyme A thiolase [Oryza sativa (japonica cultivar-group)] NP_974900 42573608 acetyl-CoA C-acyltransferase, putative/3-ketoacyl-CoA thiolase, putative [Arabidopsis thaliana] NP_974901 42573610 acetyl-CoA C-acyltransferase, putative/3-ketoacyl-CoA thiolase, putative [Arabidopsis thaliana] NP_984262 45188039 ADR165Cp [Eremothecium gossypii] XP_389497 46134945 hypothetical protein FG09321.1 [Gibberella zeae PH-1] XP_401186 49074048 hypothetical protein UM03571.1 [Ustilago maydis 521] XP_405546 49087148 hypothetical protein AN1409.2 [Aspergillus nidulans FGSC A4] XP_449306 50293789 unnamed protein product [Candida glabrata] XP_449306 50293789 unnamed protein product [Candida glabrata] XP_450298 50899020 putative acetyl-CoA C-acyltransferase [Oryza sativa (japonica cultivar- group)] XP_453599 50307241 unnamed protein product [Kluyveromyces lactis] XP_460741 50424309 unnamed protein product [Debaryomyces hansenii] XP_500646 50546253 hypothetical protein [Yarrowia lipolytica]

TABLE 8 Examples of HMG-CoA synthase polypeptides. ACCESSION GI PROTEIN DESCRIPTION B55729 1083370 hydroxymethylglutaryl-CoA synthase (EC 4.1.3.5), mitochondrial - mouse P54869 1708235 Hydroxymethylglutaryl-CoA synthase, mitochondrial precursor (HMG- CoA synthase) S13887 86312 hydroxymethylglutaryl-CoA synthase (EC 4.1.3.5) - chicken S27197 284048 hydroxymethylglutaryl-CoA synthase (EC 4.1.3.5), cytosolic, fibroblast isoform - human AAA37076 387072 3-hydroxy-3-methylglutaryl coenzyme A synthase (HMG CoA) AAF89580 9621905 3-hydroxy-3-methylglutaryl coenzyme A synthase [Dendroctonus jeffreyi] AAH00297 33991031 HMGCS1 protein [Homo sapiens] AAH31363 21618633 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 [Mus musculus] AAH42929 27552834 Hmgcs1-prov protein [Xenopus laevis] AAH79694 50925193 MGC80816 protein [Xenopus laevis] AAH83543 54035469 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 2 [Rattus norvegicus] AAO52569 28830079 similar to Homo sapiens (Human). Hypothetical protein FLJ40785 [Dictyostelium discoideum] AAP35966 30583443 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 (soluble) [Homo sapiens] BAB23657 12836439 unnamed protein product [Mus musculus] BAC04559 21754758 unnamed protein product [Homo sapiens] BAC05233 21758044 unnamed protein product [Mus musculus] CAA52032 1772495 hydroxymethylglutaryl-CoA synthase [Blattella germanica] CAC18553 11602786 putative 3-hydroxy-3-methylglutaryl coenzyme A synthase [Phycomyces blakesleeanus] CAG33131 48145817 HMGCS2 [Homo sapiens] CAH92111 55730782 hypothetical protein [Pongo pygmaeus] CAI22408 56205097 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 2 (mitochondrial) [Homo sapiens] EAK97451 46438115 hypothetical protein CaO19.7312 [Candida albicans SC5314] EAL25034 54635631 GA18098-PA [Drosophila pseudoobscura] NP_002121 54020720 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 (soluble) [Homo sapiens] NP_013580 6323509 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase [Saccharomyces cerevisiae] NP_032282 31560689 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 2 [Mus musculus] NP_058964 8393538 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 [Rattus norvegicus] NP_593859 19114771 hydroxymethylglutaryl-coa synthase (EC 4.1.3.5) [Schizosaccharomyces pombe] NP_666054 31981842 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 [Mus musculus] NP_725570 24654139 CG4311-PE, isoform E [Drosophila melanogaster] NP_775117 27465521 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 2 [Rattus norvegicus] NP_957379 41055180 similar to 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 [Danio rerio] NP_983739 45187516 ADL356Cp [Eremothecium gossypii] NP_990742 45382279 3-hydroxy-3-methylglutaryl-CoA synthase [Gallus gallus] NP_999545 47523816 hydroxymethylglutaryl-CoA synthase [Sus scrofa] XP_315872 58387870 ENSANGP00000017491 [Anopheles gambiae str. PEST] XP_323241 32405256 probable hydroxymethylglutaryl-CoA synthase [MIPS] [Neurospora crassa] XP_368218 39973655 hypothetical protein MG01026.4 [Magnaporthe grisea 70-15] XP_389442 46134253 conserved hypothetical protein [Gibberella zeae PH-1] XP_397202 48141273 similar to CG4311-PA [Apis mellifera] XP_402977 49078452 hypothetical protein UM05362.1 [Ustilago maydis 521] XP_409060 49095198 hypothetical protein AN4923.2 [Aspergillus nidulans FGSC A4] XP_446972 50289085 unnamed protein product [Candida glabrata] XP_453529 50307101 unnamed protein product [Kluyveromyces lactis] XP_456470 50405663 unnamed protein product [Debaryomyces hansenii] XP_506052 50557288 hypothetical protein [Yarrowia lipolytica] XP_513693 55587844 PREDICTED: 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 2 (mitochondrial) [Pan troglodytes] XP_536483 57085299 PREDICTED: similar to HMGCS1 protein [Canis familiaris] XP_569805 58265298 hydroxymethylglutaryl-CoA synthase, putative [Cryptococcus XP_571930 58269548 conserved hypothetical protein [Cryptococcus neoformans var. neoformans JEC21]

TABLE 9 Examples of HMG-CoA reductase polypeptides. ACCESSION GI PROTEIN DESCRIPTION A23586 90238 hydroxymethylglutaryl-CoA reductase (EC 1.1.1.88) - golden hamster O74164 11132850 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) P51639 1708252 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) P54960 1708251 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) Q12649 18276268 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) Q29512 2495262 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) Q9Y7D2 11133211 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) S30338 422383 hydroxymethylglutaryl-CoA reductase (NADPH2) (EC 1.1.1.34) - German cockroach S72194 7450066 hydroxymethylglutaryl-CoA reductase (EC 1.1.1.88) - fission yeast (Schizosaccharomyces pombe) AAA36989 387052 3-hydroxy-3-methylglutaryl coenzyme A [Mesocricetus auratus] AAA37077 305355 3-hydroxy-3-methylglutaral coenzyme A reductase (EC 1.1.1.34) AAA49740 214237 3-hydroxy-3-methylglutaryl-coenzyme A reductase AAD20975 9817458 3-hydroxy-3-methylglutaryl coenzyme A reductase [Ips paraconfusus] AAH74197 49257596 Unknown (protein for MGC:82101) [Xenopus laevis] AAL09351 15824453 3-hydroxy-3-methylglutaryl coenzyme A reductase [Ips pini] AAO85434 29468180 HMG-CoA reductase [Aspergillus nidulans] AAP72015 32165622 3-hydroxy-3-methylglutaryl-Coenzyme A reductase [Homo sapiens] AAR02862 45272118 HMG-CoA reductase [Dicentrarchus labrax] AAT92819 51013051 YLR450W [Saccharomyces cerevisiae] BAC20567 23574646 HMG-CoA reductase [Penicillium citrinum] CAA63970 4376229 HMG-CoA-reductase; hydroxymethylglutaryl-CoA reductase (NADPH) [Gibberella fujikuroi] CAE47850 41581201 3-hydroxy-3-methylglutaryl-coenzyme a reductase, putative [Aspergillus fumigatus] CAF92135 47213283 unnamed protein product [Tetraodon nigroviridis] CAH92577 55731745 hypothetical protein [Pongo pygmaeus] EAK94577 46435190 hypothetical protein CaO19.8633 [Candida albicans SC5314] EAL20195 50257490 hypothetical protein CNBF0070 [Cryptococcus neoformans var. neoformans B-3501A] AAF80374 8886086 HMG-CoA reductase [Dendroctonus jeffreyi] NP_013555 6323483 Hmg2p [Saccharomyces cerevisiae] NP_013636 6323565 Hmg1p [Saccharomyces cerevisiae] NP_032281 56119096 3-hydroxy-3-methylglutaryl-Coenzyme A reductase [Mus musculus] NP_037266 40538852 3-hydroxy-3-methylglutaryl-Coenzyme A reductase [Rattus norvegicus] NP_588235 19075735 3-hydroxy-3-methylglutaryl-coenzyme a reductase [Schizosaccharomyces pombe] NP_985010 45190756 AER152Wp [Eremothecium gossypii] NP_989816 45383193 3-hydroxy-3-methylglutaryl-CoA reductase [Gallus gallus] NP_999724 47551099 HMGCoA reductase [Strongylocentrotus purpuratus] XP_324892 32408825 hypothetical protein [Neurospora crassa] XP_364130 39955070 hypothetical protein MG08975.4 [Magnaporthe grisea 70-15] XP_389373 46134115 hypothetical protein FG09197.1 [Gibberella zeae PH-1] XP_400629 49072680 hypothetical protein UM03014.1 [Ustilago maydis 521] XP_405730 49087632 hypothetical protein AN1593.2 [Aspergillus nidulans FGSC A4] XP_407954 49092986 hypothetical protein AN3817.2 [Aspergillus nidulans FGSC A4] XP_449268 50293713 unnamed protein product [Candida glabrata] XP_451740 50303597 unnamed protein product [Kluyveromyces lactis] XP_458872 50420671 unnamed protein product [Debaryomyces hansenii] XP_503558 50552167 hypothetical protein [Yarrowia lipolytica ] XP_536323 57084803 PREDICTED: similar to 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) [Caniss ] XP_571450 58268588 hydroxymethylglutaryl-CoA reductase (NADPH), putative [Cryptococcus neoformans var. neoformans JEC21]

TABLE 10 Examples of mevalonate kinase polypeptides. ACCESSION GI PROTEIN DESCRIPTION XP_386088 46123069 hypothetical protein FG05912.1 [Gibberella zeae PH-1] XP_408006 49093090 hypothetical protein AN3869.2 [Aspergillus nidulans FGSC A4] XP_370449 39978123 hypothetical protein MG06946.4 [Magnaporthe grisea 70-15] EAL04797 46445529 hypothetical protein CaO19.4809 [Candida albicans SC5314] XP_322935 32404644 hypothetical protein ((AL513444) related to MEVALONATE KINASE [Neurospora crassa]) NP_001007 55925207 zgc:103473 [Danio rerio] XP_460851 50424525 unnamed protein product [Debaryomyces hansenii] XP_567851 58260882 cystathionine beta-lyase, putative [Cryptococcus neoformans var. neoformans JEC21] XP_567850 58260880 cystathionine beta-lyase, putative [Cryptococcus neoformans var. neoformans JEC21] AAQ02416 33303805 mevalonate kinase [synthetic construct] CAA53059 450346 unnamed protein product [Hepatitis B virus] AAH16140 16359371 MVK protein [Homo sapiens] AAH05606 13542811 Mevalonate kinase [Mus musculus] XP_403111 49078786 hypothetical protein UM05496.1 [Ustilago maydis 521] XP_452532 50305147 unnamed protein product [Kluyveromyces lactis] CAG08527 47226511 unnamed protein product [Tetraodon nigroviridis] XP_446138 50287417 unnamed protein product [Candida glabrata] AAO51522 28828936 similar to Rattus norvegicus (Rat). Mevalonate kinase (EC 2.7.1.36) (MK) [Dictyostelium discoideum] NP_985191 45190937 AER335Wp [Eremothecium gossypii] XP_500956 50546973 hypothetical protein [Yarrowia lipolytica] NP_013935 6323864 Erg12p [Saccharomyces cerevisiae] AAD45421 5578718 mevalonate kinase [Arabidopsis thaliana] NP_920723 37532842 putative mevalonate kinase [Oryza sativa (japonica cultivar-group)] NP_851084 30690651 mevalonate kinase (MK) [Arabidopsis thaliana] AAL18925 16417946 mevalonate kinase [Hevea brasiliensis] NP_788338 28573850 CG33009-PA, isoform A [Drosophila melanogaster] AAU20834 51988124 Hypothetical protein Y42G9A.4b [Caenorhabditis elegans] AAU87813 52839819 Hypothetical protein Y42G9A.4d [Caenorhabditis elegans] AAU20835 51988125 Hypothetical protein Y42G9A.4c [Caenorhabditis elegans] YP_183887 57641409 mevalonate kinase [Thermococcus kodakaraensis KOD1] NP_143478 14591399 mevalonate kinase [Pyrococcus horikoshii OT3] BAA24409 2804172 mevalonate kinase [Saccharomyces cerevisiae] NP_126232 14520757 mevalonate kinase [Pyrococcus abyssi GE5] XP_522574 55639331 PREDICTED: similar to Mevalonate kinase (MK) [Pan troglodytes] NP_071114 11499870 mevalonate kinase (mvk) [Archaeoglobus fulgidus DSM 4304] XP_423949 50797461 PREDICTED: similar to mevalonate kinase [Gallus gallus] NP_633786 21227864 Mevalonate kinase [Methanosarcina mazei Go1] ZP_002971 48840229 COG1577: Mevalonate kinase [Methanosarcina barkeri str. fusaro] EAH50787 44170778 unknown [environmental sequence] NP_615566 20089491 mevalonate kinase [Methanosarcina acetivorans C2A] 1VISA 40890012 Chain A, Crystal Structure Of Mevalonate Kinase EAK03559 44549994 unknown [environmental sequence] NP_248080 15669275 mevalonate kinase [Methanocaldococcus jannaschii DSM 2661] 1KKHA 20150886 Chain A, Crystal Structure Of The Methanococcus Jannaschii Mevalonate Kinase Q50559 2497518 Mevalonate kinase (MK) CAF88123 47200914 unnamed protein product [Tetraodon nigroviridis] NP_275189 15678075 mevalonate kinase [Methanothermobacter thermautotrophicus str. Delta H] EAI88745 44383877 unknown [environmental sequence] ZP_002040 46141948 COG1577: Mevalonate kinase [Methanococcoides burtonii DSM 6242] XP_543435 57105916 PREDICTED: similar to Mevalonate kinase (MK) [Canis familiaris] EAI38920 44313360 unknown [environmental sequence] NP_148611 14602065 mevalonate kinase [Aeropyrum pernix K1] EAD08953 43286228 unknown [environmental sequence] EAD45697 43361720 unknown [environmental sequence] YP_134862 55377012 mevalonate kinase [Haloarcula marismortui ATCC 43049] NP_720650 24378695 putative mevalonate kinase [Streptococcus mutans UA159] NP_614276 20094429 Mevalonate kinase [Methanopyrus kandleri AV19] E84270 25409931 mevalonate kinase [imported] - Halobacterium sp. NRC-1 NP_691146 23097680 mevalonate kinase [Oceanobacillus iheyensis HTE831] ZP_003233 48870579 COG1577: Mevalonate kinase [Pediococcus pentosaceus ATCC 25745] AAG02440 9937386 mevalonate kinase [Enterococcus faecalis] EAD12278 43292898 unknown [environmental sequence] NP_498328 17555862 mevalonate kinase (64.1 kD) (3H214) [Caenorhabditis elegans] EAB31483 42928976 unknown [environmental sequence] ZP_003319 50590618 COG1577: Mevalonate kinase [Streptococcus suis 89/1591] NP_814642 29375488 mevalonate kinase [Enterococcus faecalis V583] AC1434 25514495 mevalonate kinase homolog lin0010 [imported] - Listeria innocua (strain Clip11262) ZP_003577 53796847 COG1577: Mevalonate kinase [Chloroflexus aurantiacus] EAD82048 43454743 unknown [environmental sequence] CAE73618 39586491 Hypothetical protein CBG21109 [Caenorhabditis briggsae] YP_012624 46906235 mevalonate kinase [Listeria monocytogenes str. 4b F2365] NP_988455 45358898 Mevalonate kinase [Methanococcus maripaludis S2] ZP_002348 47097293 mevalonate kinase [Listeria monocytogenes str. 1/2a F6854] ZP_002862 48824993 COG1577: Mevalonate kinase [Enterococcus faecium] ZP_002307 47093020 mevalonate kinase [Listeria monocytogenes str. 4b H7858] NP_597102 19173299 MEVALONATE KINASE [Encephalitozoon cuniculi] CAD24422 20429111 phosphomevalonate kinase [Paracoccus zeaxanthinifaciens] NP_785308 28378416 mevalonate kinase [Lactobacillus plantarum WCFS1] EAA39098 29247539 GLP_305_18405_19430 [Giardia lamblia ATCC 50803] NP_819638 29653946 phosphomevalonate kinase, putative [Coxiella burnetii RSA 493] EAH49746 44168765 unknown [environmental sequence] EAH49745 44168764 unknown [environmental sequence] NP_378182 15922513 hypothetical protein ST2185 [Sulfolobus tokodaii str. 7] ZP_000459 23002259 COG1577: Mevalonate kinase [Lactobacillus gasseri] H90181 25393827 mevalonate kinase [imported] - Sulfolobus solfataricus YP_054120 50405028 Mevalonate kinase, putative [Paramecium tetraurelia] BAB07790 9695270 mevalonate kinase [Streptomyces sp. CL190] AAG02435 9937379 mevalonate kinase [Staphylococcus epidermidis] NP_560495 18313828 mevalonate kinase [Pyrobaculum aerophilum str. IM2] YP_187834 57866187 mevalonate kinase [Staphylococcus epidermidis RP62A] EAK40782 44602942 unknown [environmental sequence] CAC51370 15212070 mevalonate kinase [Lactobacillus helveticus] AAG02424 9937364 mevalonate kinase [Staphylococcus aureus] YP_185521 57651465 mevalonate kinase [Staphylococcus aureus subsp. aureus COL] YP_040044 49482820 mevalonate kinase [Staphylococcus aureus subsp. aureus MRSA252] YP_194037 58337452 mevalonate kinase [Lactobacillus acidophilus NCFM] D86675 25400965 mevalonate kinase [imported] - Lactococcus lactis subsp. lactis (strain IL1403) NP_763916 27467279 mevalonate kinase [Staphylococcus epidermidis ATCC 12228] CAF89434 47197810 unnamed protein product [Tetraodon nigroviridis] EAF38333 43767792 unknown [environmental sequence] EAK46841 44611394 unknown [environmental sequence] H89827 25507776 mevalonate kinase [imported] - Staphylococcus aureus (strain N315) ZP_003149 48861061 COG0153: Galactokinase [Microbulbifer degradans 2-40] EAK17824 44570143 unknown [environmental sequence] EAH86276 44235719 unknown [environmental sequence] YP_118418 54024176 putative mevalonate kinase [Nocardia farcinica IFM 10152] ZP_003196 48865749 COG1577: Mevalonate kinase [Oenococcus oeni PSU-1] AAG02430 9937372 mevalonate kinase [Staphylococcus haemolyticus] NP_269075 15674901 mevalonate kinase [Streptococcus pyogenes M1 GAS] NP_802520 28896170 putative mevalonate kinase [Streptococcus pyogenes SSI-1] AAL97579 19748102 mevalonate kinase [Streptococcus pyogenes MGAS8232] ZP_003666 56808907 COG1577: Mevalonate kinase [Streptococcus pyogenes M49 591] NP_965060 42519130 mevalonate kinase [Lactobacillus johnsonii NCC 533] NP_819639 29653947 mevalonate kinase, putative [Coxiella burnetii RSA 493] EAD97024 43484567 unknown [environmental sequence] BAD86800 57753870 mevalonate kinase [Streptomyces sp. KO-3988]

TABLE 11 Examples of phosphomevalonate kinase polypeptides. ACCESSION GI PROTEIN DESCRIPTION AAA34596 171479 phosphomevalonate kinase XP_452514 50305111 unnamed protein product [Kluyveromyces lactis] NP_985210 45190956 AER354Wp [Eremothecium gossypii] XP_446144 50287429 unnamed protein product [Candida glabrata] XP_462340 50427455 unnamed protein product [Debaryomyces hansenii] EAL04096 46444824 hypothetical protein CaO19.12076 [Candida albicans SC5314] EAL03941 46444668 hypothetical protein CaO19.4606 [Candida albicans SC5314] XP_503619 50552418 hypothetical protein [Yarrowia lipolytica] XP_389940 46136497 hypothetical protein FG09764.1 [Gibberella zeae PH-1] XP_329795 32418634 hypothetical protein [Neurospora crassa] XP_369652 39976529 hypothetical protein MG05812.4 [Magnaporthe grisea 70-15] XP_406448 49089559 hypothetical protein AN2311.2 [Aspergillus nidulans FGSC A4] NP_593421 19114333 putative phosphomevalonate kinase [Schizosaccharomyces pombe] XP_568385 58261950 expressed protein [Cryptococcus neoformans var. neoformans JEC21] EAL17628 50254887 hypothetical protein CNBM0120 [Cryptococcus neoformans var. neoformans B-3501A] AAL18926 16417948 phosphomevalonate kinase [Hevea brasiliensis] BAD43274 51969164 hypothetical protein [Arabidopsis thaliana] BAD44652 51971975 hypothetical protein [Arabidopsis thaliana] XP_398375 49068172 hypothetical protein UM00760.1 [Ustilago maydis 521] BAD44486 51971643 hypothetical protein [Arabidopsis thaliana] F90479 25393214 (Phospho) mevalonate kinase, probable [imported] - Sulfolobus solfataricus YP_194039 58337454 phosphomevalonate kinase [Lactobacillus acidophilus NCFM]

TABLE 12 Examples of mevalonate pyrophosphate decarboxylase polypeptides. ACCESSION GI PROTEIN DESCRIPTION AAT93171 51013755 YNR043W [Saccharomyces cerevisiae] 1FI4A 13786942 Chain A, The X-Ray Crystal Structure Of Mevalonate 5-Diphosphate Decarboxylase At 2.3 Angstrom Resolution. XP_455548 50311049 unnamed protein product [Kluyveromyces lactis] XP_445335 50285813 unnamed protein product [Candida glabrata] XP_456912 50409853 unnamed protein product [Debaryomyces hansenii] NP_986435 45200865 AGL232Cp [Eremothecium gossypii] AAF19399 6625790 diphosphomevalonate decarboxylase MVD1 [Candida albicans] XP_328845 32416734 hypothetical protein [Neurospora crassa] XP_505041 50555265 hypothetical protein [Yarrowia lipolytica] NP_594027 19114939 diphosphomevalonate decarboxylase [Schizosaccharomyces pombe] XP_364905 39963452 hypothetical protein MG09750.4 [Magnaporthe grisea 70-15] XP_390600 46137817 hypothetical protein FG10424.1 [Gibberella zeae PH-1] XP_408551 49094180 hypothetical protein AN4414.2 [Aspergillus nidulans FGSC A4] AAA34506 7544604 ORF [Saccharomyces cerevisiae] EAL18927 50256200 hypothetical protein CNBI1880 [Cryptococcus neoformans var. neoformans B-3501A] XP_568247 58261674 diphosphomevalonate decarboxylase, putative [Cryptococcus neoformans var. neoformans JEC21] XP_402794 49077992 hypothetical protein UM05179.1 [Ustilago maydis 521] AAH81784 51980639 Mevalonate (diphospho) decarboxylase [Rattus norvegicus] EAL00166 46440864 hypothetical protein CaO19.6105 [Candida albicans SC5314] NP_619597 20149736 mevalonate (diphospho) decarboxylase [Mus musculus] NP_112324 13592005 mevalonate (diphospho) decarboxylase [Rattus norvegicus] BAC40852 26354448 unnamed protein product [Mus musculus] XP_546783 57087071 Mevalonate pyrophosphate decarboxylase [Canis familiaris] Q99JF5 23814095 Diphosphomevalonate decarboxylase (Mevalonate pyrophosphate decarboxylase) AAH63907 39645379 LOC394871 protein [Xenopus tropicalis] CAF99534 47230341 unnamed protein product [Tetraodon nigroviridis] AAP35576 30582699 mevalonate (diphospho) decarboxylase [Homo sapiens] AAP36301 30584105 Homo sapiens mevalonate (diphospho) decarboxylase [synthetic construct] AAL18927 16417950 mevalonate disphosphate decarboxylase [Hevea brasiliensis] AAV32433 54292590 mevalonate disphosphate decarboxylase [Ginkgo biloba] AAP68208 31711704 At2g38700 [Arabidopsis thaliana] AAM64988 21593039 mevalonate diphosphate decarboxylase [Arabidopsis thaliana] NP_566995 18410026 mevalonate diphosphate decarboxylase, putative [Arabidopsis thaliana] XP_423130 50771155 PREDICTED: similar to mevalonate pyrophosphate decarboxylase [Gallus gallus] AAM65192 21593243 diphosphomevalonate decarboxylase-like protein [Arabidopsis thaliana] NP_001007 55925435 zgc:100824 [Danio rerio] NP_573068 28571205 CG8239-PA [Drosophila melanogaster] BAD27942 50252009 putative mevalonate disphosphate decarboxylase [Oryza sativa (japonica cultivar-group)] T47584 11281655 diphosphomevalonate decarboxylase (EC 4.1.1.33) F24B22.210 [similarity] - Arabidopsis thaliana XP_307373 31196851 ENSANGP00000013738 [Anopheles gambiae str. PEST] CAE73245 39591192 Hypothetical protein CBG20661 [Caenorhabditis briggsae] NP_496966 17537201 mevalonate decarboxylase (41.5 kD) (2O512) [Caenorhabditis elegans] XP_393230 48121058 similar to mevalonate pyrophosphate decarboxylase [Apis mellifera] G90479 25393662 diphosphomevalonate decarboxylase, probable [imported] - Sulfolobus solfataricus NP_496967 17537203 mevalonate decarboxylase (2O512) [Caenorhabditis elegans] NP_691147 23097681 mevalonate diphosphate decarboxylase [Oceanobacillus iheyensis HTE831] EAL29282 54640164 GA20922-PA [Drosophila pseudoobscura] AD1434 25515042 mevalonate diphosphate decarboxylase homolog lin0011 [imported] - Listeria innocua (strain Clip11262) ZP_002308 47093021 diphosphomevalonate decarboxylase [Listeria monocytogenes str. 4b H7858] YP_012625 46906236 diphosphomevalonate decarboxylase [Listeria monocytogenes str. 4b F2365] ZP_002348 47097294 diphosphomevalonate decarboxylase [Listeria monocytogenes str. 1/2a F6854] NP_819637 29653945 diphosphomevalonate decarboxylase [Coxiella burnetii RSA 493] NP_376888 15921219 hypothetical diphosphomevalonate decarboxylase [Sulfolobus tokodaii str. 7] ZP_003319 50590617 COG3407: Mevalonate pyrophosphate decarboxylase [Streptococcus suis 89/1591] NP_585805 19074299 MEVALONATE PYROPHOSPHATE DECARBOXYLASE [Encephalitozoon cuniculi] YP_187835 57866188 mevalonate diphosphate decarboxylase [Staphylococcus epidermidis RP62A] CAD24423 20429112 mevalonate diphosphate decarboxylase [Paracoccus zeaxanthinifaciens] AAG02431 9937373 mevalonate diphosphate decarboxylase [Staphylococcus haemolyticus] NP_763917 27467280 mevalonate diphosphate decarboxylase [Staphylococcus epidermidis ATCC 12228] AAG02446 9937394 mevalonate diphosphate decarboxylase [Enterococcus faecium] ZP_002863 48824994 COG3407: Mevalonate pyrophosphate decarboxylase [Enterococcus faecium] AAG02441 9937387 mevalonate diphosphate decarboxylase [Enterococcus faecalis] YP_185522 57651466 mevalonate diphosphate decarboxylase [Staphylococcus aureus subsp. aureus COL] A89828 25505863 mevalonate diphosphate decarboxylase [imported] - Staphylococcus aureus (strain N315) NP_814641 29375487 mevalonate diphosphate decarboxylase [Enterococcus faecalis V583] YP_040045 49482821 mevalonate diphosphate decarboxylase [Staphylococcus aureus subsp. aureus MRSA252] NP_785307 28378415 diphosphomevalonate decarboxylase [Lactobacillus plantarum WCFS1] ZP_003196 48865750 COG3407: Mevalonate pyrophosphate decarboxylase [Oenococcus oeni PSU-1] ZP_003233 48870580 COG3407: Mevalonate pyrophosphate decarboxylase [Pediococcus pentosaceus ATCC 25745] E86675 25400967 diphosphomevalonate decarboxylase [imported] - Lactococcus lactis subsp. lactis (strain IL1403) EAE31110 43552684 unknown [environmental sequence] BAB07791 9695271 diphosphomevalonate decarboxylase [Streptomyces sp. CL190] CAC51371 15212071 mevalonate diphosphate decarboxylase [Lactobacillus helveticus] ZP_000459 23002258 COG3407: Mevalonate pyrophosphate decarboxylase [Lactobacillus gasseri] NP_965061 42519131 mevalonate pyrophosphate decarboxylase [Lactobacillus johnsonii NCC 533] BAD86801 57753871 mevalonate diphosphate decarboxylase [Streptomyces sp. KO-3988] YP_194038 58337453 mevalonate diphosphate decarboxylase [Lactobacillus acidophilus NCFM] YP_118419 54024177 putative diphosphomevalonate decarboxylase [Nocardia farcinica IFM 10152] EAK18820 44571499 unknown [environmental sequence] EAI85935 44379784 unknown [environmental sequence] NP_721336 24379381 putative mevalonate diphosphate decarboxylase [Streptococcus mutans UA159] D95044 25388338 diphosphomevalonate decarboxylase [imported] - Streptococcus pneumoniae (strain TIGR4) AAG02456 9937408 mevalonate diphosphate decarboxylase [Streptococcus pneumoniae] C97914 25511486 diphosphomevalonate decarboxylase (EC 4.1.1.33) [imported] - Streptococcus pneumoniae (strain R6) EAK47683 44612560 unknown [environmental sequence] EAB86425 43039778 unknown [environmental sequence] YP_140971 55822530 mevalonate pyrophosphate decarboxylase [Streptococcus thermophilus CNRZ1066] YP_139081 55820639 mevalonate pyrophosphate decarboxylase [Streptococcus thermophilus LMG 18311] BAD07376 40882372 mevalonate diphosphate decarboxylase [Actinoplanes sp. A40644] NP_968512 42523132 Diphosphomevalonate decarboxylase [Bdellovibrio bacteriovorus HD100] EAI06705 44265427 unknown [environmental sequence] YP_060018 50914046 Diphosphomevalonate decarboxylase [Streptococcus pyogenes MGAS10394] AAG02451 9937401 mevalonate diphosphate decarboxylase [Streptococcus pyogenes] NP_269076 15674902 mevalonate pyrophosphate decarboxylase [Streptococcus pyogenes M1 GAS] ZP_003666 56808906 COG3407: Mevalonate pyrophosphate decarboxylase [Streptococcus pyogenes M49 591] NP_688323 22537472 diphosphomevalonate decarboxylase [Streptococcus agalactiae 2603V/R] NP_735832 25011437 hypothetical protein gbs1395 [Streptococcus agalactiae NEM316] EAC40267 43149093 unknown [environmental sequence] AAL97580 19748103 mevalonate pyrophosphate decarboxylase [Streptococcus pyogenes MGAS8232] EAI76915 44367119 unknown [environmental sequence] EAD35042 43339207 unknown [environmental sequence] YP_073129 51598941 mevalonate pyrophosphate decarboxylase [Borrelia garinii PBi] EAI90092 44385501 unknown [environmental sequence] BAB07818 9711347 mevalonate diphosphate decaroboxylase [Kitasatospora griseola] EAD72850 43433025 unknown [environmental sequence] NP_212820 15595031 mevalonate pyrophosphate decarboxylase [Borrelia burgdorferi B31] YP_124337 54297968 hypothetical protein lpp2023 [Legionella pneumophila str. Paris] YP_096056 52842257 mevalonate diphosphate decarboxylase [Legionella pneumophila subsp. pneumophila str. Philadelphia 1] EAA39903 29248368 GLP_479_14176_13169 [Giardia lamblia ATCC 50803] EAH06252 44088237 unknown [environmental sequence] YP_127354 54294939 hypothetical protein lpl2018 [Legionella pneumophila str. Lens] EAD45753 43361830 unknown [environmental sequence] NP_802519 28896169 putative mevalonate pyrophosphate decarboxylase [Streptococcus pyogenes SSI-1]

TABLE 13 Examples of IPP isomerase polypeptides. ACCESSION GI PROTEIN DESCRIPTION NP_015208 6325140 Idi1p [Saccharomyces cerevisiae] XP_448008 50291151 unnamed protein product [Candida glabrata] NP_983828 45187605 ADL268Cp [Eremothecium gossypii] XP_455121 50310203 unnamed protein product [Kluyveromyces lactis] XP_462358 50427491 unnamed protein product [Debaryomyces hansenii] EAL01685 46442395 potential isopentenyl diphosphate isomerase [Candida albicans SC5314] XP_504974 50555131 hypothetical protein [Yarrowia lipolytica] XP_328425 32415894 hypothetical protein [Neurospora crassa] XP_367200 39971619 hypothetical protein MG07125.4 [Magnaporthe grisea 70-15] XP_389898 46136413 conserved hypothetical protein [Gibberella zeae PH-1] XP_404716 49085144 hypothetical protein AN0579.2 [Aspergillus nidulans FGSC A4] CAD37150 21627818 isopentenyl-diphosphate delta-isomerase [Aspergillus fumigatus] NP_595164 19111956 isopentenyl-diphosphate delta-isomerase [Schizosaccharomyces pombe] XP_566641 58258457 isopentenyl-diphosphate delta-isomerase, putative [Cryptococcus neoformans var. neoformans JEC21] XP_402453 49077100 hypothetical protein UM04838.1 [Ustilago maydis 521] O35586 6225528 Isopentenyl-diphosphate delta-isomerase 1 (IPP isomerase 1) AAP36609 30584713 Homo sapiens isopentenyl-diphosphate delta isomerase [synthetic construct] AAF37873 7188790 isopentenyl pyrophosphate isomerase [Dictyostelium discoideum] NP_445991 16758306 isopentenyl-diphosphate delta isomerase [Rattus norvegicus] O42641 6225529 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) BAA33979 3790386 IPP isomerase [Xanthophyllomyces dendrorhous] Q13907 6225527 Isopentenyl-diphosphate delta-isomerase 1 (IPP isomerase 1) AAH22418 48257241 IDI1 protein [Homo sapiens] AAH19227 48257312 IDI1 protein [Homo sapiens] AAH57827 35505325 IDI1 protein [Homo sapiens] NP_004499 40018633 isopentenyl-diphosphate delta isomerase [Homo sapiens] AAH89786 58477715 Unknown (protein for MGC:108635) [Rattus norvegicus] CAH91844 55730243 hypothetical protein [Pongo pygmaeus] XP_418561 50732281 PREDICTED: similar to isopentenyl-diphosphate delta isomerase; IPP isomerase [Gallus gallus] AAH06999 48257093 IDI1 protein [Homo sapiens] CAF98782 47225155 unnamed protein product [Tetraodon nigroviridis] NP_808875 29366820 isopentenyl-diphosphate delta isomerase [Mus musculus] XP_507622 55633353 PREDICTED: similar to isopentenyl-diphosphate delta isomerase; IPP isomerase [Pan troglodytes] AAH82648 52139082 LOC494671 protein [Xenopus laevis] NP_001011 58332496 hypothetical LOC496783 [Xenopus tropicalis] AAF29976 6856556 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Lactuca sativa] AAG10423 9971806 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Tagetes erecta] O48964 6225525 Isopentenyl-diphosphate delta-isomerase I (IPP isomerase I) (Isopentenyl pyrophosphate isomerase I) AAF29973 6856550 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Adonis palaestina] AAF29977 6856558 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Tagetes erecta] AAQ84167 35186998 isopentenyl pyrophosphate isomerase [Pueraria montana var. lobata] AAF29974 6856552 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Adonis palaestina] Q39472 6225526 Isopentenyl-diphosphate delta-isomerase I (IPP isomerase I) (Isopentenyl pyrophosphate isomerase I) S49588 1085973 isopentenyl-diphosphate Delta-isomerase (EC 5.3.3.2) 1 - Clarkia breweri (fragment) AAL91980 19568939 isopentenyl pyrophosphate isomerase IDI1 [Melaleuca alternifolia] BAB40973 13603406 isopentenyl diphosphate isomerase 1 [Nicotiana tabacum] AAF29975 6856554 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Lactuca sativa] T52027 25493162 isopentenyl-diphosphate Delta-isomerase (EC 5.3.3.2) 1 [validated] - Haematococcus pluvialis AAL91979 19568937 isopentenyl pyrophosphate isomerase IDI2 [Melaleuca alternifolia] T46812 11362218 isopentenyl-diphosphate Delta-isomerase (EC 5.3.3.2) [imported] - Haematococcus pluvialis T51248 11362217 isopentenyl-diphosphate Delta-isomerase (EC 5.3.3.2) 2 [validated] - Haematococcus pluvialis BAB40974 13603408 isopentenyl diphosphate isomerase 2 [Nicotiana tabacum] O48965 6225532 Isopentenyl-diphosphate delta-isomerase II (IPP isomerase II) (Isopentenyl pyrophosphate isomerase II) XP_225509 34877710 similar to isopentenyl diphosphate delta-isomerase type 2 [Rattus norvegicus] XP_506401 51963472 PREDICTED OJ1612_A04.101 gene product [Oryza sativa (japonica cultivar-group)] AAF29978 6856560 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Oryza sativa] AAH76541 50369278 Unknown (protein for IMAGE:7037641) [Danio rerio] AAT94033 51038230 putative isopentenyl-diphosphate delta-isomerase [Oryza sativa (japonica cultivar-group)] XP_225502 34876517 similar to isopentenyl diphosphate delta-isomerase type 2 [Rattus norvegicus] Q39471 6225533 Isopentenyl-diphosphate delta-isomerase II (IPP isomerase II) (Isopentenyl pyrophosphate isomerase II) AAB67743 1213450 isopentenyl pyrophosphate isomerase [Clarkia breweri] NP_197148 22326844 isopentenyl-diphosphate delta-isomerase I [Arabidopsis thaliana] BAB09611 9759126 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Arabidopsis thaliana] AAD41766 5305669 isopentenyl pyrophosphate isomerase [Hevea brasiliensis] AAB67741 1213442 isopentenyl pyrophosphate isomerase [Arabidopsis thaliana] XP_395125 48101420 similar to CG8646-PA [Apis mellifera] AAN28784 23505849 At3g02780/F13E7_28 [Arabidopsis thaliana] AAF36996 7110585 isopentenyl pyrophosphate:dimethylallyl pyrophosphate isomerase [Brassica oleracea var. botrytis] BAB16690 15289752 putative IPP isomerase [Eucommia ulmoides] AAQ14869 33340598 isopentenyl pyrophosphate isomerase [Zea mays] BAC65421 28971819 isopentenyl-diphosphate delta-isomerase [Periploca sepium] S71369 2129625 isopentenyl-diphosphate Delta-isomerase (EC 5.3.3.2) 1 - Arabidopsis thaliana AAF29979 6856562 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Arabidopsis thaliana] AAF29980 6856564 isopentenyl pyrophosphate:dimethyllallyl pyrophosphate isomerase [Arabidopsis thaliana] AAP21674 30267831 isopentenyl diphosphate delta-isomerase type 2 [Mus musculus] Q39664 6225534 Isopentenyl-diphosphate delta-isomerase II (IPP isomerase II) (Isopentenyl pyrophosphate isomerase II) NP_650962 24648688 CG5919-PA [Drosophila melanogaster] AAM50284 21429130 RE22306p [Drosophila melanogaster] XP_321388 58395620 ENSANGP00000011643 [Anopheles gambiae str. PEST] Q9BXS1 20978506 Isopentenyl-diphosphate delta-isomerase 2 (IPP isomerase 2)) T07979 7484383 isopentenyl-diphosphate Delta-isomerase (EC 5.3.3.2) ipi1 - Chlamydomonas reinhardtii XP_225508 34876527 similar to diphosphate dimethylallyl diphosphate isomerase 2 [Rattus norvegicus] AAT92102 51011386 isopentenyl-diphosphate delta-isomerase [Ixodes pacificus] XP_225507 34876555 similar to isopentenyl diphosphate delta-isomerase type 2 [Rattus norvegicus] XP_344623 34876537 similar to isopentenyl diphosphate delta-isomerase type 2 [Rattus norvegicus] S44843 630677 K06H7.3 protein - Caenorhabditis elegans XP_225498 27687955 similar to diphosphate dimethylallyl diphosphate isomerase 2 [Rattus norvegicus] AAT08468 47013849 isopentenyl-diphosphate isomerase [Caenorhabditis elegans] EAI79636 44370808 unknown [environmental sequence] CAE75055 39587401 Hypothetical protein CBG22969 [Caenorhabditis briggsae] EAL04047 46444775 potential isopentenyl diphosphate isomerase fragment [Candida albicans SC5314] XP_225528 34876543 similar to isopentenyl diphosphate delta-isomerase type 2 [Rattus norvegicus] XP_544282 57040602 PREDICTED: similar to isopentenyl-diphosphate delta isomerase [Canis familiaris] XP_225511 27688013 similar to diphosphate dimethylallyl diphosphate isomerase 2 [Rattus norvegicus] P26173 114853 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Bacteriochlorophyll synthase 20 kDa chain) EAJ04069 44405322 unknown [environmental sequence] EAH27496 44127513 unknown [environmental sequence] AAF91499 9653280 isopentenyl pyrophosphate isomerase [Daucus carota] AAM48661 21328655 isopentyl-diphosphate delta-isomerase [uncultured proteobacterium] EAK17826 44570145 unknown [environmental sequence] EAD59515 43391069 unknown [environmental sequence] YP_128702 54307682 hypothetical isopentenyldiphosphate isomerase [Photobacterium profundum SS9] EAK66656 44639203 unknown [environmental sequence] YP_118189 54023947 putative isopentenyldiphosphate isomerase [Nocardia farcinica IFM 10152] T50740 11282665 isopentenyl diphosphate isomerase homolog [imported] - Rhodobacter sphaeroides ZP_002077 46193541 COG1443: Isopentenyldiphosphate isomerase [Rhodobacter sphaeroides 2.4.1] EAK16470 44568229 unknown [environmental sequence] YP_165403 56695056 isopentyl-diphosphate delta-isomerase [Silicibacter pomeroyi DSS-3] EAD08775 43285885 unknown [environmental sequence] YP_195623 58616494 putative isopentenyl-diphosphate delta-isomerase [Azoarcus sp. EbN1] EAI38918 44313358 unknown [environmental sequence] NP_930583 37527239 hypothetical protein plu3365 [Photorhabdus luminescens subsp. laumondii TTO1] YP_160254 56478665 isopentenyl-diphosphate delta-isomerase [Azoarcus sp. EbN1] EAH69842 44206571 unknown [environmental sequence] EAK26254 44582307 unknown [environmental sequence] AAR24381 38569721 isopentyl-diphosphate delta isomerase [Sus scrofa] AAM48607 21328600 isopentenyl-diphosphate delta-isomerase, putative [uncultured proteobacterium] EAD82049 43454744 unknown [environmental sequence] ZP_001924 45914126 COG1443: Isopentenyldiphosphate isomerase [Mesorhizobium sp. BNC1] YP_056780 50843553 isopentenyl-diphosphate delta-isomerase [Propionibacterium acnes KPA171202] YP_050880 50121713 putative isopentenyl-diphosphate delta-isomerase [Erwinia carotovora subsp. atroseptica SCRI1043] EAF29235 43749645 unknown [environmental sequence] NP_630823 21225044 putative IPP isomerase [Streptomyces coelicolor A3(2)] Q82MJ7 34582349 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) ZP_003374 52010110 COG1443: Isopentenyldiphosphate isomerase [Silicibacter sp. TM1040] AAS75819 45737905 isopentenyl/dimethylallyl diphosphate isomerase [Nicotiana benthamiana] Q8KP37 30913023 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) XP_507621 55633351 PREDICTED: similar to isopentenyl-diphosphate delta isomerase 2 [Pan troglodytes] XP_344621 34876521 similar to diphosphate dimethylallyl diphosphate isomerase 2 [Rattus norvegicus] XP_346322 34880719 similar to diphosphate dimethylallyl diphosphate isomerase 2 [Rattus norvegicus] YP_152060 56414985 probable isomerase [Salmonella enterica subsp. enterica serovar Paratypi A str. ATCC 9150] AAT42442 48429280 putative isopentenyl diphosphate isomerase [Edwardsiella ictaluri] Q9KK75 13878536 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) NP_806649 29143307 probable isomerase [Salmonella enterica subsp. enterica serovar Typhi Ty2] YP_063124 50955836 isopentenyldiphosphate isomerase [Leifsonia xyli subsp. xyli str. CTCB07] Q8FND7 46395593 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) CAF20647 41326485 ISOPENTENYLDIPHOSPHATE ISOMERASE [Corynebacterium glutamicum ATCC 13032] Q8NN99 23821718 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) Q7X5H2 46395586 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) NP_336246 15841209 isopentenyl-diphosphate delta-isomerase [Mycobacterium tuberculosis CDC1551] Q83MJ9 46395588 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) P60923 46395576 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) Q8FE75 31563050 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) (Isopentenyl pyrophosphate isomerase) 1R67A 38493022 Chain A, Y104a Mutant Of E. Coli Ipp Isomerase Q9KWD1 13878537 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) Q7VEU0 46395585 Isopentenyl-diphosphate delta-isomerase (IPP isomerase) B84333 25410326 isopentenyl pyrophosphate isomerase [imported] - Halobacterium sp. NRC-1 NP_417365 16130791 isopentenyl diphosphate isomerase [Escherichia coli K12] E85944 25355426 probable enzyme Z4227 [imported] - Escherichia coli (strain O157:H7, substrain EDL933) 1HZTA 15826050 Chain A, Crystal Structure Of Metal-Free Isopentenyl Diphosphate:dimethylallyl Diphosphate Isomerase 1PVFB 50513321 Chain B, E. Coli Ipp Isomerase In Complex With Diphosphate EAD63579 43403471 unknown [environmental sequence] 1I9AB 13786886 Chain B, Mevalonate 5-Diphosphate Decarboxylase And Isopentenyl Diphosphate Isomerase YP_012992 46906603 MutT/nudix family protein [Listeria monocytogenes str. 4b F2365] ZP_002293 47091503 MutT/nudix family protein [Listeria monocytogenes str. 4b H7858] EAI37194 44310821 unknown [environmental sequence] YP_137864 55380014 probable isopentenyl-diphosphate delta-isomerase [Haloarcula marismortui ATCC 43049] CAD92056 42516867 putative isopentenyl diphosphate isomerase type 1 [Haloferax mediterranei] 1OW2B 42543244 Chain B, Isopentenylpyrophosphate-Dimethylallylpyrophosphate Isomerase: Complex Of C67a Mutant With Eipp

TABLE 14 Examples of FPP synthase polypeptides. ACCESSION GI PROTEIN DESCRIPTION Q92250 2497455 Farnesyl pyrophosphate synthetase (FPP synthetase) (FPS) (Farnesyl diphosphate synthetase) XP_363065 39948036 hypothetical protein MG08649.4 [Magnaporthe grisea 70-15] XP_386960 46124813 FPPS_GIBFU Farnesyl pyrophosphate synthetase (FPP synthetase) (FPS) [Gibberella zeae PH-1] Q92235 3122099 Farnesyl pyrophosphate synthetase (FPP synthetase) (FPS) (Farnesyl diphosphate synthetase) XP_412149 49116518 hypothetical protein AN8012.2 [Aspergillus nidulans FGSC A4] XP_503599 50552378 hypothetical protein [Yarrowia lipolytica] NP_593299 19114211 farnesyl pyrophosphate synthetase [Schizosaccharomyces pombe] CAD42869 21955860 farnesyl pyrophosphate synthase [Mucor circinelloides f. lusitanicus] XP_448787 50292709 unnamed protein product [Candida glabrata] NP_012368 6322294 Erg20p [Saccharomyces cerevisiae] T42081 7433997 farnesyl-pyrophosphate synthetase - fission yeast (Schizosaccharomyces pombe) (fragment) EAK93751 46434339 likely farnesyl diphosphate synthetase [Candida albicans SC5314] XP_451300 50302727 FPPS_KLULA [Kluyveromyces lactis] XP_571137 58267962 isoprenoid biosynthesis-related protein, putative [Cryptococcus neoformans var. neoformans JEC21] XP_460720 50424267 unnamed protein product [Debaryomyces hansenii] NP_984739 45190485 AEL122Wp [Eremothecium gossypii] BAD15361 46367743 farnesyl diphosphate synthase [Lactarius chrysorrheus] S71433 7433991 farnesyl-pyrophosphate synthetase - ergot fungus (fragment) CAA65643 1523990 farnesyl pyrophosphate synthetase [Claviceps purpurea] XP_399061 49069544 hypothetical protein UM01446.1 [Ustilago maydis 521] S71432 7433990 farnesyl-pyrophosphate synthetase - Sphaceloma manihoticola (fragment) AAH68912 46249832 MGC83119 protein [Xenopus laevis] 1FPS 1065289 Chain, Avian Farnesyl Diphosphate Synthase (Fps) (E.C.2.5.1.10) P08836 3915686 Farnesyl pyrophosphate synthetase (FPP synthetase) (FPS) (Farnesyl diphosphate synthetase) AAH83515 53733369 Unknown (protein for IMAGE:7049076) [Danio rerio] 1UBX 1942050 Chain, Structure Of Farnesyl Pyrophosphate Synthetase 1UBY 1942051 Chain, Structure Of Farnesyl Pyrophosphate Synthetase AAF37872 7188788 farnesyl diphosphate synthase [Dictyostelium discoideum] NP_803463 29135293 farnesyl diphosphate synthase [Bos taurus] AAK63847 14488053 farnesyl diphosphate synthase [Mentha × piperita] AAV58896 55710092 farnesyl diphosphate synthase [Centella asiatica] T06272 7433988 farnesyl-pyrophosphate synthetase FPS1 - tomato JC4846 2117737 farnesyl-pyrophosphate synthetase - Artemisia annua P05369 120478 Farnesyl pyrophosphate synthetase (FPP synthetase) (FPS) (Farnesyl diphosphate synthetase) O24241 25452945 Farnesyl pyrophosphate synthetase 1 (FPP synthetase 1) O24242 25452946 Farnesyl pyrophosphate synthetase 2 (FPP synthetase 2) AAH59125 37590777 Testis-specific farnesyl pyrophosphate synthetase [Rattus norvegicus] AAH48497 28913418 Farnesyl diphosphate synthetase [Mus musculus] AAP74720 32329199 farnesyl diphosphate synthase [Artemisia tridentata subsp. spiciformis] CAG11850 47225367 unnamed protein product [Tetraodon nigroviridis] AAM51429 21436457 putative farnesyl-pyrophosphate synthetase FPS2 [Arabidopsis thaliana] AAP74719 32329197 farnesyl diphosphate synthase [Artemisia tridentata subsp. spiciformis] AAM08927 20135548 farnesyl pyrophosphate synthase [Malus × domestica] XP_537252 57089113 PREDICTED: similar to farnesyl diphosphate synthase [Canis familiaris] AAQ56011 34013692 farnesyl diphosphate synthase [Hevea brasiliensis] AAQ14872 33340604 truncated geranylgeranyl-diphosphate synthase [Zea mays] AAQ14871 33340602 geranylgeranyl-diphosphate synthase [Zea mays] AAD17204 4324960 farnesyl diphosphate synthase [Artemisia annua] AAH87886 56789674 Farnesyl diphosphate synthetase [Mus musculus] AAK68152 14573639 farnesyldiphosphate synthase [×Citrofortunella microcarpa] AAA52423 182399 farnesyl pyrophosphate synthetase (EC 2.5.1.1) S66470 2129849 farnesyl-pyrophosphate synthetase fps1 - white lupine CAA29064 4725 unnamed protein product [Saccharomyces cerevisiae] CAI12715 55957735 farnesyl diphosphate synthase [Homo sapiens] BAA03523 40788949 KIAA1293 [Homo sapiens] P14324 1346031 Farnesyl pyrophosphate synthetase (FPP synthetase) S66471 2129850 farnesyl-pyrophosphate synthetase fps2 - white lupine AAA35820 182405 farnesyl pyrophosphate synthetase CAA59170 1491641 dimethylallyltransferase [Capsicum annuum] BAB16687 15289750 putative FPP synthase 1 [Eucommia ulmoides] CAA72793 1922251 farnesyl pyrophosphate synthase [Gossypium arboreum] CAH91070 55728661 hypothetical protein [Pongo pygmaeus] AAK58594 14279425 farnesyl pyrophosphate synthase [Humulus lupulus] AAB07264 1146159 farnesyl diphosphate synthase short form [Arabidopsis thaliana] Q09152 21431776 Farnesyl pyrophosphate synthetase 1, mitochondrial precursor (FPP synthetase 1) O64905 6016044 Farnesyl pyrophosphate synthetase (FPP synthetase) BAB60822 14422406 putative FPP synthase 2 [Eucommia ulmoides] S52009 1076319 farnesyl-pyrophosphate synthetase FPS1 - Arabidopsis thaliana NP_917118 34911542 putative farnesyl-pyrophosphate synthetase [Oryza sativa (japonica cultivar-group)] AAD32648 4894899 farnesyl diphosphate synthase [Artemisia annua] AAA40960 203582 cholesterol-regulated protein CR39 AAR27053 38684029 farnesyl diphosphate synthetase [Ginkgo biloba] AAU43998 52353430 putative farnesyl pyrophosphate synthase [Oryza sativa (japonica cultivar-group)] AAL82595 18958450 farnesyl pyrophosphare synthase [Musa acuminata] NP_917069 34911444 putative farnesyl-pyrophosphate synthetase [Oryza sativa (japonica cultivar-group)] XP_228802 34879769 similar to testis-specific farnesyl pyrophosphate synthetase [Rattus norvegicus] BAD81810 56785155 putative farnesyl-pyrophosphate synthetase fps2 [Oryza sativa (japonica cultivar-group)] AAN62522 24796660 farnesyl pyrophosphate synthetase [Eucommia ulmoides] NP_595334 19112126 farnesyl pyrophosphate synthetase [Schizosaccharomyces pombe] T52066 25458583 probable farnesyl pyrophosphate synthase [imported] - rice AAL49067 17946048 RE52884p [Drosophila melanogaster] CAA08919 3395483 dimethylallyltransferase; farnesyl pyrophosphate synthase [Drosophila melanogaster] XP_547662 57089869 PREDICTED: similar to farnesyl diphosphate synthase [Canis familiaris] EAL26135 54636732 GA11601-PA [Drosophila pseudoobscura] BAB60821 14422404 putative FPP synthase 1 [Eucommia ulmoides] AAP74721 32329201 chrysanthemyl diphosphate synthase [Artemisia tridentata subsp. spiciformis] XP_496902 51466663 PREDICTED: similar to Farnesyl pyrophosphate synthetase (FPP synthetase) [Homo sapiens] XP_474182 50929309 OSJNBa0071I13.18 [Oryza sativa (japonica cultivar-group)] CAA87327 1160178 partial sequence [Homo sapiens] BAD20729 47776234 farnesyl pyrophosphate synthase [Candida glabrata] BAC53873 30984142 farnesyl pyrophosphate synthase [Phaseolus lunatus] BAB69490 15991313 farnesyl pyrophosphate syntase [Bombyx mori] NP_974565 42572937 farnesyl pyrophosphate synthetase 2 (FPS2)/FPP synthetase 2 [Arabidopsis thaliana] CAA08918 5678609 dimethylallyltransferase; farnesyl pyrophosphate synthase [Agrotis ipsilon] AAP86267 32527731 Ac2-125 [Rattus norvegicus] AAO17735 30522953 farnesyl pyrophosphate synthase [Trypanosoma brucei] AAK71861 14647139 farnesyl pyrophosphate synthase [Trypanosoma cruzi] AAL73357 18478919 farnesyl diphosphate synthase precursor [Trypanosoma cruzi] AAO63552 29124957 putative farnesyl pyrophosphate synthase [Plasmodium falciparum] CAI00471 56498227 farnesyl pyrophosphate synthase, putative [Plasmodium berghei] NP_701155 23508486 farnesyl pyrophosphate synthase, putative [Plasmodium falciparum 3D7] XP_474180 50929305 OSJNBa0071I13.16 [Oryza sativa (japonica cultivar-group)] AAL73358 18478922 farnesyl diphosphate synthase precursor [Trypanosoma cruzi] EAH48995 44167328 unknown [environmental sequence] NP_493027 17508563 farnesyl pyrophosphate synthetase (1M510) [Caenorhabditis elegans] CAE71711 39580204 Hypothetical protein CBG18688 [Caenorhabditis briggsae] XP_487220 51766977 similar to farnesyl pyrophosphate synthase [Mus musculus]

TABLE 15 Examples of GGPP synthase polypeptides. ACCESSION GI PROTEIN DESCRIPTION AAT92871 51013155 YPL069C [Saccharomyces cerevisiae] XP_447025 50289191 unnamed protein product [Candida glabrata] NP_984623 45190369 AEL238Cp [Eremothecium gossypii] XP_390273 46137163 GGPP_GIBFU Geranylgeranyl pyrophosphate synthetase (GGPP synthetase) [Gibberella zeae PH-1] XP_404791 49085320 hypothetical protein AN0654.2 [Aspergillus nidulans FGSC A4] XP_368486 39974191 hypothetical protein MG00758.4 [Magnaporthe grisea 70-15] Q92236 6831550 Geranylgeranyl pyrophosphate synthetase (GGPP synthetase) AAO85432 29468176 geranylgeranyl diphosphate synthase [Aspergillus nidulans] XP_572774 58271236 farnesyltranstransferase, putative [Cryptococcus neoformans var. neoformans JEC21] XP_502923 50550901 hypothetical protein [Yarrowia lipolytica] AAK11525 13021716 geranylgeranyl pyrophosphate synthase [Penicillium paxilli] XP_326920 32412880 GERANYLGERANYL PYROPHOSPHATE SYNTHETASE (GGPP SYNTHETASE) [Neurospora crassa] CAF32032 42820719 geranylgeranyl pyrophosphate synthetase, putative [Aspergillus fumigatus] BAD29965 50355599 Phoma betae geranylgeranyl diphosphate synthase XP_384767 46117498 hypothetical protein FG04591.1 [Gibberella zeae PH-1] BAD29970 50355631 geranylgeranyldiphosphate synthase [Phoma betae] CAB89115 7649674 geranylgeranyl pyrophosphate synthase [Mucor circinelloides f. lusitanicus] CAG09545 47229030 unnamed protein product [Tetraodon nigroviridis] CAI13753 55960163 geranylgeranyl diphosphate synthase 1 [Homo sapiens] AAH69913 47124116 Geranylgeranyl diphosphate synthase 1 [Mus musculus] AAH67768 45709211 GGPS1 protein [Homo sapiens] XP_455003 50309979 unnamed protein product [Kluyveromyces lactis] P56966 9296978 Geranylgeranyl pyrophosphate synthetase (GGPP synthetase) NP_001007 56090562 geranylgeranyl diphosphate synthase 1 [Rattus norvegicus] AAT65717 49409613 geranylgeranyl diphosphate synthase [Aspergillus flavus] NP_956329 41053321 geranylgeranyl diphosphate synthase 1 [Danio rerio] BAA90525 6899844 geranylgeranyl diphosphate synthase [Nigrospora sphaerica] XP_405729 49087630 hypothetical protein AN1592.2 [Aspergillus nidulans FGSC A4] AAK11531 13021724 geranylgeranyl pyrophosphate synthase [Penicillium paxilli] XP_412280 49119197 hypothetical protein AN8143.2 [Aspergillus nidulans FGSC A4] AAC05273 2944400 geranylgeranyl pyrophosphate synthase [Drosophila melanogaster] NP_523958 24660002 CG8593-PA [Drosophila melanogaster] XP_402074 49076128 hypothetical protein UM04459.1 [Ustilago maydis 521] EAL30191 54641441 GA21189-PA [Drosophila pseudoobscura] XP_536340 57084951 PREDICTED: hypothetical protein XP_536340 [Canis familiaris] XP_424685 50811194 PREDICTED: similar to geranylgeranyl diphosphate synthase 1, partial [Gallus gallus] AAH06798 13905030 Ggps1 protein [Mus musculus] AAP06018 29841005 similar to NM_010282 geranylgeranyl diphosphate synthase 1; GGPP synthase [Schistosoma japonicum] XP_460338 50423511 unnamed protein product [Debaryomyces hansenii] AAC05595 2957271 geranylgeranyl pyrophosphate synthase [Drosophila melanogaster] EAK92197 46432727 hypothetical protein CaO19.6674 [Candida albicans SC5314] XP_535573 57108760 PREDICTED: similar to geranylgeranyl diphosphate synthase 1 [Canis familiaris] AAH83212 53734594 Zgc:56514 protein [Danio rerio] XP_486466 51827552 similar to geranylgeranyl diphosphate synthase 1; GGPP synthase [Mus musculus] CAH18006 51469024 geranylgeranyl diphosphate synthase [Fusarium proliferatum] CAA75568 3549881 geranylgeranyl diphosphate synthase [Gibberella fujikuroi] XP_397455 48143654 similar to CG8593-PA [Apis mellifera] XP_410947 49101294 hypothetical protein AN6810.2 [Aspergillus nidulans FGSC A4] XP_381914 46109712 hypothetical protein FG01738.1 [Gibberella zeae PH-1] XP_364478 39959279 hypothetical protein MG09448.4 [Magnaporthe grisea 70-15] XP_360889 39942704 hypothetical protein MG03432.4 [Magnaporthe grisea 70-15] XP_369218 39975655 hypothetical protein MG00026.4 [Magnaporthe grisea 70-15] XP_406544 49089926 hypothetical protein AN2407.2 [Aspergillus nidulans FGSC A4] XP_367595 39972409 hypothetical protein MG07506.4 [Magnaporthe grisea 70-15] XP_363775 39952117 hypothetical protein MG01701.4 [Magnaporthe grisea 70-15] XP_368486 39974191 hypothetical protein MG00758.4 [Magnaporthe grisea 70-15] XP_390273 46137163 GGPP_GIBFU Geranylgeranyl pyrophosphate synthetase (GGPP synthetase) [Gibberella zeae PH-1] Q92236 6831550 Geranylgeranyl pyrophosphate synthetase (GGPP synthetase) AAK11525 13021716 geranylgeranyl pyrophosphate synthase [Penicillium paxilli] CAF32032 42820719 geranylgeranyl pyrophosphate synthetase, putative [Aspergillus fumigatus] XP_404791 49085320 hypothetical protein AN0654.2 [Aspergillus nidulans FGSC A4] AAO85432 29468176 geranylgeranyl diphosphate synthase [Aspergillus nidulans] BAD29965 50355599 Phoma betae geranylgeranyl diphosphate synthase BAD29970 50355631 geranylgeranyldiphosphate synthase [Phoma betae] BAA90525 6899844 geranylgeranyl diphosphate synthase [Nigrospora sphaerica] AAT65717 49409613 geranylgeranyl diphosphate synthase [Aspergillus flavus] XP_384767 46117498 hypothetical protein FG04591.1 [Gibberella zeae PH-1] CAB89115 7649674 geranylgeranyl pyrophosphate synthase [Mucor circinelloides f. lusitanicus] XP_572774 58271236 farnesyltranstransferase, putative [Cryptococcus neoformans var. neoformans JEC21] AAK11531 13021724 geranylgeranyl pyrophosphate synthase [Penicillium paxilli] XP_502923 50550901 hypothetical protein [Yarrowia lipolytica] CAI13753 55960163 geranylgeranyl diphosphate synthase 1 [Homo sapiens] CAG09545 47229030 unnamed protein product [Tetraodon nigroviridis] XP_412280 49119197 hypothetical protein AN8143.2 [Aspergillus nidulans FGSC A4] P56966 9296978 Geranylgeranyl pyrophosphate synthetase (GGPP synthetase) NP_001007 56090562 geranylgeranyl diphosphate synthase 1 [Rattus norvegicus] AAH69913 47124116 Geranylgeranyl diphosphate synthase 1 [Mus musculus] AAH67768 45709211 GGPS1 protein [Homo sapiens] NP_956329 41053321 geranylgeranyl diphosphate synthase 1 [Danio rerio] EAL30191 54641441 GA21189-PA [Drosophila pseudoobscura] XP_424685 50811194 PREDICTED: similar to geranylgeranyl diphosphate synthase 1, partial [Gallus gallus] XP_536340 57084951 PREDICTED: hypothetical protein XP_536340 [Canis familiaris] NP_523958 24660002 CG8593-PA [Drosophila melanogaster] AAC05273 2944400 geranylgeranyl pyrophosphate synthase [Drosophila melanogaster] XP_405729 49087630 hypothetical protein AN1592.2 [Aspergillus nidulans FGSC A4] AAC05595 2957271 geranylgeranyl pyrophosphate synthase [Drosophila melanogaster] XP_402074 49076128 hypothetical protein UM04459.1 [Ustilago maydis 521] AAP06018 29841005 similar to NM_010282 geranylgeranyl diphosphate synthase 1; GGPP synthase [Schistosoma japonicum] AAH06798 13905030 Ggps1 protein [Mus musculus] XP_535573 57108760 PREDICTED: similar to geranylgeranyl diphosphate synthase 1 [Canis familiaris] AAH83212 53734594 Zgc: 56514 protein [Danio rerio] AAP21085 30097620 albino-3 [Neurospora crassa] NP_984623 45190369 AEL238Cp [Eremothecium gossypii] XP_447025 50289191 unnamed protein product [Candida glabrata] AAT92871 51013155 YPL069C [Saccharomyces cerevisiae] XP_486466 51827552 similar to geranylgeranyl diphosphate synthase 1; GGPP synthase [Mus musculus] XP_410947 49101294 hypothetical protein AN6810.2 [Aspergillus nidulans FGSC A4] XP_397455 48143654 similar to CG8593-PA [Apis mellifera] XP_455003 50309979 unnamed protein product [Kluyveromyces lactis] EAK92197 46432727 hypothetical protein CaO19.6674 [Candida albicans SC5314] XP_381914 46109712 hypothetical protein FG01738.1 [Gibberella zeae PH-1] XP_460338 50423511 unnamed protein product [Debaryomyces hansenii] CAH18006 51469024 geranylgeranyl diphosphate synthase [Fusarium proliferatum] XP_360889 39942704 hypothetical protein MG03432.4 [Magnaporthe grisea 70-15] XP_406544 49089926 hypothetical protein AN2407.2 [Aspergillus nidulans FGSC A4] XP_364478 39959279 hypothetical protein MG09448.4 [Magnaporthe grisea 70-15] XP_363775 39952117 hypothetical protein MG01701.4 [Magnaporthe grisea 70-15] XP_367595 39972409 hypothetical protein MG07506.4 [Magnaporthe grisea 70-15] XP_369218 39975655 hypothetical protein MG00026.4 [Magnaporthe grisea 70-15] C39273 483124 phytoene synthase - Erwinia herbicola BAB79600 18143445 crtE [Pantoea agglomerans pv. milletiae] BAA14124 216682 crtE [Pantoea ananatis] AAN85596 27228290 Geranylgeranyl Pyrophosphate Synthase [Pantoea stewartii] AAA32797 413730 geranylgeranyl pyrophosphate synthase Q08291 585326 Geranyltranstransferase (Farnesyl-diphosphate synthase) (FPP synthase) S52584 1073293 crtE protein - Erwinia herbicola S53722 1076576 farnesyltranstransferase (EC 2.5.1.29) precursor - pepper AAC44848 1842242 geranylgeranyl synthase BAA19583 1944371 geranylgeranyl pyrophosphate synthase [Arabidopsis thaliana] S71230 2129674 geranylgeranyl pyrophosphate synthase (EC 2.5.1.—) 2 precursor - Arabidopsis thaliana BAA23157 2578822 geranyl geranyl pyrophosphate synthase [Arabidopsis thaliana] AAC77874 3885426 geranylgeranyl pyrophosphate synthase [Helianthus annuus] CAB38744 4490594 geranylgeranyl pyrophosphate synthase [Rhodobacter sphaeroides] BAA78047 4958920 GGPP synthase [Daucus carota] BAA82613 5631295 geranylgeranyl diphosphate synthase (SelGGPS) [Synechococcus elongatus] CAB56064 5912297 geranylgeranyl pyrophosphate synthase [Paracoccus marcusii] BAA86284 6277254 geranylgeranyl pyrophosphate synthase [Croton sublyratus] T11021 7447356 farnesyltranstransferase (EC 2.5.1.29) - white lupine AAF78199 8650415 geranylgeranyl synthase [Bradyrhizobium sp. ORS278] AAG10424 9971808 GGDP synthase [Tagetes erecta] CAC10561 10637876 gpp synthase large subunit [Mentha × piperita] T50879 11279298 phytoene synthase [imported] - Rubrivivax gelatinosus BAB01343 11994221 geranyl geranyl pyrophosphate synthase-like protein [Arabidopsis thaliana] Q42698 13431546 Geranylgeranyl pyrophosphate synthetase, chloroplast precursor (GGPP synthetase) Q43133 13431547 Geranylgeranyl pyrophosphate synthetase, chloroplast precursor (GGPP synthetase) P54976 13878921 Geranylgeranyl pyrophosphate synthetase (GGPP synthetase) (Farnesyltranstransferase) BAB50600 14023995 geranyltranstransferase; farnesyl-diphosphate synthase [Mesorhizobium loti MAFF303099] BAB60678 14325238 geranylgeranyl diphosphate synthase [Hevea brasiliensis] BAB60820 14422402 putative GGPP synthase [Eucommia ulmoides] NP_189589 15228704 geranylgeranyl pyrophosphate synthase, putative/GGPP synthetase [Arabidopsis thaliana] NP_188651 15231055 geranylgeranyl pyrophosphate synthase, putative/GGPP synthetase [Arabidopsis thaliana] NP_188069 15231869 geranylgeranyl pyrophosphate synthase, putative/GGPP synthetase [Arabidopsis thaliana] NP_188073 15231881 geranylgeranyl pyrophosphate synthase, putative/GGPP synthetase [Arabidopsis thaliana] AAL01997 15553715 farnesyl diphosphate synthase [Xanthobacter sp. Py2] AAL01998 15553717 geranylgeranyl diphosphate synthase [Xanthobacter sp. Py2] NP_252732 15599238 geranyltranstransferase [Pseudomonas aeruginosa PAO1] NP_245470 15602398 IspA [Pasteurella multocida subsp. multocida str. Pm70] NP_390308 16079484 hypothetical protein BSU24280 [Bacillus subtilis subsp. subtilis str. 168] NP_440010 16329282 geranylgeranyl pyrophosphate synthase [Synechocystis sp. PCC 6803] NP_440010 16329282 geranylgeranyl pyrophosphate synthase [Synechocystis sp. PCC 6803] AAL17614 17352451 geranylgeranyl diphosphate synthase [Abies grandis] NP_520343 17546941 PROBABLE GERANYLTRANSTRANSFERASE (FARNESYL- DIPHOSPHATE SYNTHASE) PROTEIN [Ralstonia solanacearum GM AAL76349 18645048 geranyltranstransferase [uncultured proteobacterium] AAM21638 20386366 geranylgeranyl pyrophosphate synthase [Cistus incanus subsp. creticus] AAM21639 20386368 geranylgeranyl pyrophosphate synthase [Cistus incanus subsp. creticus] NP_622916 20807745 Geranylgeranyl pyrophosphate synthase [Thermoanaerobacter tengcongensis MB4] AAM48650 21328644 geranylgeranyl pyrophosphate synthetase [uncultured proteobacterium] NP_659794 21492720 probable polyprenyl synthetase. [Rhizobium etli] AAM64496 21592547 putative geranylgeranyl pyrophosphate synthase GGPS3 [Arabidopsis thaliana] AAM65107 21593158 geranylgeranyl pyrophosphate synthase [Arabidopsis thaliana] NP_680811 22297564 geranylgeranyl pyrophosphate synthase [Thermosynechococcus elongatus BP-1] ZP_000474 23003800 COG0142: Geranylgeranyl pyrophosphate synthase [Lactobacillus gasseri] ZP_001252 23469933 COG0142: Geranylgeranyl pyrophosphate synthase [Pseudomonas syringae pv. syringae B728a] NP_698760 23502633 geranyltranstransferase [Brucella suis 1330] E84566 25313373 probable geranylgeranyl pyrophosphate synthase [imported] - Arabidopsis thaliana F85434 25313385 geranylgeranyl pyrophosphate synthase [imported] - Arabidopsis thaliana AC1245 25313389 geranyltranstransferase homolog lmo1363 [imported] - Listeria monocytogenes (strain EGD-e) E83997 25313393 geranyltranstransferase BH2781 [imported] - Bacillus halodurans (strain C-125) G84566 25313395 probable geranylgeranyl pyrophosphate synthase [imported] - Arabidopsis thaliana AH2910 25315863 geranyltranstransferase [imported] - Agrobacterium tumefaciens (strain C58, Dupont) D87505 25398795 geranyltranstransferase [imported] - Caulobacter crescentus A89932 25505949 hypothetical protein ispA [imported] - Staphylococcus aureus (strain N315) F97685 25520741 geranyltransferase (AF203881) [imported] - Agrobacterium tumefaciens (strain C58, Cereon) AI3285 25527013 geranyltranstransferase (EC 2.5.1.10) [imported] - Brucella melitensis (strain 16M) BAC42571 26450928 putative geranylgeranyl pyrophosphate synthase GGPS3 [Arabidopsis thaliana] NP_785195 28378303 geranyltranstransferase [Lactobacillus plantarum WCFS1] NP_790546 28867927 geranyltranstransferase [Pseudomonas syringae pv. tomato str. DC3000] AAO63392 28950937 At2g23800 [Arabidopsis thaliana] AAO93113 29893480 geranylgeranyl pyrophosphate synthetase; CrtE [Rubrivivax gelatinosus] NP_833891 30022260 Dimethylallyltransferase [Bacillus cereus ATCC 14579] AAP59037 31621279 CrtE [Thiocapsa roseopersicina] ZP_001374 32039216 COG0142: Geranylgeranyl pyrophosphate synthase [Pseudomonas aeruginosa UCBPP-PA14] NP_864766 32471772 geranylgeranyl pyrophosphate synthetase [precursor] [Rhodopirellula baltica SH 1] NP_875521 33240579 Geranylgeranyl pyrophosphate synthase [Prochlorococcus marinus subsp. marinus str. CCMP1375] NP_881399 33593755 probable geranyltranstransferase [Bordetella pertussis Tohama I] NP_884694 33597051 probable geranyltranstransferase [Bordetella parapertussis 12822] NP_888456 33600896 probable geranyltranstransferase [Bordetella bronchiseptica RB50] NP_893187 33861626 Polyprenyl synthetase [Prochlorococcus marinus subsp. pastoris str. CCMP1986] NP_894940 33863380 Polyprenyl synthetase [Prochlorococcus marinus str. MIT 9313] NP_896835 33865276 geranylgeranyl pyrophosphate synthase [Synechococcus sp. WH 8102] NP_896835 33865276 geranylgeranyl pyrophosphate synthase [Synechococcus sp. WH 8102] AAQ65086 34365549 At3g14530 [Arabidopsis thaliana] NP_945877 39933601 putative geranyltranstransferase (farnesyl-diphosphate synthase) [Rhodopseudomonas palustris CGA009] NP_946867 39934591 geranylgeranyl pyrophosphate synthase [Rhodopseudomonas palustris CGA009] NP_952815 39996864 geranyltranstransferase [Geobacter sulfurreducens PCA] AAR37805 40062934 polyprenyl synthetase [uncultured bacterium 442] AAR37858 40062988 geranylgeranyl pyrophosphate synthetase [uncultured bacterium 443] AAR98495 41018904 geranyl geranyl synthase [Bradyrhizobium sp. ORS278] AAR99082 41059107 geranylgeranyl pyrophosphate synthase [Plectranthus barbatus] NP_965349 42519419 geranyltranstransferase [Lactobacillus johnsonii NCC 533] NP_980544 42783297 geranyltranstransferase [Bacillus cereus ATCC 10987] EAA96348 42858148 unknown [environmental sequence] EAB36506 42939031 unknown [environmental sequence] EAB36642 42939300 unknown [environmental sequence] EAC39208 43146996 unknown [environmental sequence] EAD26007 43320598 unknown [environmental sequence] EAE43084 43576643 unknown [environmental sequence] EAE70061 43630884 unknown [environmental sequence] EAF70308 43832107 unknown [environmental sequence] EAG88494 44055952 unknown [environmental sequence] EAH52060 44173220 unknown [environmental sequence] EAH78354 44221788 unknown [environmental sequence] EAH84117 44231960 unknown [environmental sequence] EAI11762 44272832 unknown [environmental sequence] EAI49391 44328289 unknown [environmental sequence] EAI54846 44336042 unknown [environmental sequence] EAI68356 44355138 unknown [environmental sequence] EAI68713 44355672 unknown [environmental sequence] EAI69401 44356609 unknown [environmental sequence] EAI73873 44362658 unknown [environmental sequence] EAJ73634 44506168 unknown [environmental sequence] EAJ77351 44511694 unknown [environmental sequence] EAK70639 44644254 unknown [environmental sequence] ZP_001751 45523854 COG0142: Geranylgeranyl pyrophosphate synthase [Crocosphaera watsonii WH 8501] AAS76253 45752710 At1g49530 [Arabidopsis thaliana] ZP_001957 45916757 COG0142: Geranylgeranyl pyrophosphate synthase [Mesorhizobium sp. BNC1] 1RTRB 46015556 Chain B, Crystal Structure Of S. Aureus Farnesyl Pyrophosphate Synthase ZP_001863 46105954 COG0142: Geranylgeranyl pyrophosphate synthase [Rubrobacter xylanophilus DSM 9941] ZP_002002 46107045 COG0142: Geranylgeranyl pyrophosphate synthase [Rubrobacter xylanophilus DSM 9941] ZP_001711 46132567 COG0142: Geranylgeranyl pyrophosphate synthase [Ralstonia eutropha JMP134] ZP_002073 46192680 COG0142: Geranylgeranyl pyrophosphate synthase [Rhodobacter sphaeroides 2.4.1] ZP_002074 46192861 COG0142: Geranylgeranyl pyrophosphate synthase [Rhodobacter sphaeroides 2.4.1] AAS82860 46241274 geranyl diphosphate synthase large subunit [Antirrhinum majus] ZP_002108 46308696 COG0142: Geranylgeranyl pyrophosphate synthase [Ehrlichia canis str. Jake] YP_010568 46579760 geranylgeranyl diphosphate synthase [Desulfovibrio vulgaris subsp. vulgaris str. Hildenborough] BAD18313 47076770 geranyltranstransferase [Geobacillus stearothermophilus] ZP_002315 47093750 geranyltranstransferase [Listeria monocytogenes str. 4b H7858] ZP_002335 47095946 geranyltranstransferase [Listeria monocytogenes str. 1/2a F6854] AAT35222 47531118 fusion of carotene synthesis proteins [synthetic construct] ZP_002401 47569437 geranyltranstransferase [Bacillus cereus G9241] ZP_002435 47573473 COG0142: Geranylgeranyl pyrophosphate synthase [Rubrivivax gelatinosus PM1] ZP_002626 48728941 COG0142: Geranylgeranyl pyrophosphate synthase [Pseudomonas fluorescens PfO-1] ZP_002702 48765678 COG0142: Geranylgeranyl pyrophosphate synthase [Rhodospirillum rubrum] ZP_002705 48766028 COG0142: Geranylgeranyl pyrophosphate synthase [Rhodospirillum rubrum] ZP_002732 48768894 COG0142: Geranylgeranyl pyrophosphate synthase [Ralstonia metallidurans CH34] ZP_002914 48834438 COG0142: Geranylgeranyl pyrophosphate synthase [Magnetococcus sp. MC-1] ZP_003024 48848203 COG0142: Geranylgeranyl pyrophosphate synthase [Novosphingobium aromaticivorans DSM 12444] ZP_003129 48858958 COG0142: Geranylgeranyl pyrophosphate synthase [Clostridium thermocellum ATCC 27405] ZP_003177 48863841 COG0142: Geranylgeranyl pyrophosphate synthase [Microbulbifer degradans 2-40] ZP_003225 48869790 COG0142: Geranylgeranyl pyrophosphate synthase [Pediococcus pentosaceus ATCC 25745] AAT51323 49086036 PA4043 [synthetic construct] ZP_003301 49236117 COG0142: Geranylgeranyl pyrophosphate synthase [Moorella thermoacetica ATCC 39073] YP_034222 49476181 Geranyltranstransferase (farnesyl-diphosphate synthase) [Bartonella henselae str. Houston-1] YP_040995 49483771 putative geranyltranstransferase [Staphylococcus aureus subsp. aureus MRSA252] YP_043579 49486358 putative geranyltranstransferase [Staphylococcus aureus subsp. aureus MSSA476] AAT71982 50253560 At3g32040 [Arabidopsis thaliana] AAT90315 50952782 putative geranylgeranyl pyrophosphate synthetase [uncultured proteobacterium eBACred25D05] YP_066435 51246551 probable geranylgeranyl pyrophosphate synthase [Desulfotalea psychrophila LSv54] YP_075673 51892982 geranylgeranyl pyrophosphate synthase [Symbiobacterium thermophilum IAM 14863] YP_085511 52141318 geranyltranstransferase [Bacillus cereus ZK] YP_092166 52786337 YqiD [Bacillus licheniformis ATCC 14580] ZP_001298 53691368 COG0142: Geranylgeranyl pyrophosphate synthase [Desulfovibrio desulfuricans G20] YP_105136 53716444 geranyltranstransferase [Burkholderia mallei ATCC 23344] YP_111769 53722784 geranyltranstransferase [Burkholderia pseudomallei K96243] ZP_003630 54030933 COG0142: Geranylgeranyl pyrophosphate synthase [Polaromonas sp. JS666] YP_129021 54308001 putative geranyltranstransferase [Photobacterium profundum SS9] AAV74395 56122554 geranylgeranyl diphosphate synthase [Adonis palaestina] AAV74396 56122556 geranylgeranyl diphosphate synthase [Adonis palaestina] YP_148246 56420928 geranyltranstransferase (farnesyl-diphosphate synthase) [Geobacillus kaustophilus HTA426] YP_156518 56461237 Geranylgeranyl pyrophosphate synthase [Idiomarina loihiensis L2TR] YP_162590 56551751 geranyltranstransferase [Zymomonas mobilis subsp. mobilis ZM4] YP_171470 56750769 geranylgeranyl pyrophosphate synthase [Synechococcus elongatus PCC 6301] YP_175959 56964228 geranyltranstransferase [Bacillus clausii KSM-K16] YP_186407 57650478 geranyltranstransferase [Staphylococcus aureus subsp. aureus COL] YP_190690 58038726 Dimethylallyltransferase [Gluconobacter oxydans 621H] AAW66658 58201026 geranyl diphosphate synthase [Picrorhiza kurrooa] YP_194187 58337602 geranylgeranyl diphosphate (GGPP) synthase [Lactobacillus acidophilus NCFM] YP_197469 58579257 Possible geranyltranstransferase [Ehrlichia ruminantium str. Welgevonden] YP_201938 58582922 geranyltranstransferase; farnesyl-diphosphate synthase [Xanthomonas oryzae pv. oryzae KACC10331] YP_196510 58617311 Possible geranyltranstransferase [Ehrlichia ruminantium str. Gardel]

TABLE 16 Examples of squalene synthetase polypeptides. ACCESSION GI PROTEIN DESCRIPTION AAA34597 171481 squalene synthetase CAA42583 3686 farnesyl-diphosphate farnesyltransferase [Saccharomyces cerevisiae] Q9HGZ6 51704336 Farnesyl-diphosphate farnesyltransferase (Squalene synthetase) (SQS) (SS) (FPP:FPP farnesyltransferase) BAB12207 9955387 squalene synthase [Candida glabrata] XP_453457 50306959 unnamed protein product [Kluyveromyces lactis] Q752X9 51701405 Farnesyl-diphosphate farnesyltransferase (Squalene synthetase) (SQS) (SS) (FPP:FPP farnesyltransferase) O74165 51701378 Farnesyl-diphosphate farnesyltransferase (Squalene synthetase) (SQS) (SS) (FPP:FPP farnesyltransferase) XP_458579 50420093 unnamed protein product [Debaryomyces hansenii] EAK95451 46436082 hypothetical protein CaO19.11099 [Candida albicans SC5314] P78589 2499979 Farnesyl-diphosphate farnesyltransferase (Squalene synthetase) (SQS) (SS) (FPP:FPP farnesyltransferase) Q9Y753 51701459 Farnesyl-diphosphate farnesyltransferase (Squalene synthetase) (SQS) (SS) (FPP:FPP farnesyltransferase) XP_407513 49092104 hypothetical protein AN3376.2 [Aspergillus nidulans FGSC A4] XP_364394 39958237 hypothetical protein MG09239.4 [Magnaporthe grisea 70-15] Q7S4Z6 51701416 Probable farnesyl-diphosphate farnesyltransferase (Squalene synthetase) CAD60581 27764301 unnamed protein product [Podospora anserina] XP_389557 46135731 hypothetical protein FG09381.1 [Gibberella zeae PH-1] NP_595363 19112155 farnesyl-diphosphate farnesyltransferase [Schizosaccharomyces pombe] B48057 477750 farnesyl-diphosphate farnesyltransferase (EC 2.5.1.21) - fission yeast (Schizosaccharomyces pombe ) NP_034321 34328173 farnesyl diphosphate farnesyl transferase 1 [Mus musculus] CAH92517 55731622 hypothetical protein [Pongo pygmaeus] AAF00038 6002565 squalene synthase [Mus musculus] P53798 1706773 Farnesyl-diphosphate farnesyltransferase (Squalene synthetase) (SQS) (SS) (FPP:FPP farnesyltransferase) NP_004453 31542632 farnesyl-diphosphate farnesyltransferase 1 [Homo sapiens] AAP36671 30584837 Homo sapiens farnesyl-diphosphate farnesyltransferase 1 [synthetic construct] 1EZFC 11514497 Chain C, Crystal Structure Of Human Squalene Synthase AAH09251 14328083 Farnesyl-diphosphate farnesyltransferase 1 [Homo sapiens] AAH84016 54035372 LOC494973 protein [Xenopus laevis] I52090 2136196 squalene synthase - human XP_420043 50745256 PREDICTED: similar to Farnesyl-diphosphate farnesyltransferase 1 [Gallus gallus] AAH81810 51858605 Farnesyl diphosphate farnesyl transferase 1 [Rattus norvegicus] CAE48363 50841455 TPA: FDFT1 protein [Bos taurus] XP_569783 58265254 farnesyl-diphosphate farnesyltransferase, putative [Cryptococcus neoformans var. neoformans JEC21] XP_569782 58265252 farnesyl-diphosphate farnesyltransferase, putative [Cryptococcus neoformans var. neoformans JEC21] XP_534557 57105080 PREDICTED: similar to FDFT1 protein [Canis familiaris] XP_401989 49075920 FDFT_USTMA Farnesyl-diphosphate farnesyltransferase (Squalene synthetase) [Ustilago maydis 521]

TABLE 17a Examples of phytoene dehydrogenase polypeptides. ACCESSION PROTEIN DESCRIPTION 1613414B crtI gene 1613414F crtD gene 1904206A methoxyneurosporene dehydrogenase 2121278A zeta carotene desaturase A86203 hypothetical protein [imported] - Arabidopsis thaliana A96612 hypothetical protein F12K22.18 [imported] - Arabidopsis thaliana A99470 phytoene dehydrogenase (desaturase) (crtI) [imported] - Sulfolobus solfataricus AAA24820 phytoene dehydrogenase [Pantoea agglomerans] AAA34001 phytoene desaturase AAA50313 photoprotective pigment AAA64981 phytoene dehydrogenase [Pantoea agglomerans] AAA91161 zeta-carotene desaturase precursor AAA99519 phytoene desaturase AAC44798 hydroxyneurosporene and rhodopin dehydrogenase [Rubrivivax gelatinosus] AAC44850 phytoene desaturase AAC48983 phytoene dehydrogenase precursor AAF78201 phytoene desaturase [Bradyrhizobium sp. ORS278] AAG10426 phytoene desaturase [Tagetes erecta] AAG14399 zeta-carotene desaturase precursor [Oryza sativa] AAG28700 CrtI [Streptomyces griseus] AAG50743 hypothetical protein [Arabidopsis thaliana] AAH85048 Unknown (protein for MGC: 97898) [Xenopus laevis] AAK51545 phytoene desaturase [Citrus × paradisi] AAK51557 zeta-carotene desaturase precursor [Citrus × paradisi] AAK64299 phytoene desaturase [Corynebacterium glutamicum] AAL02000 phytoene dehydrogenase [Xanthobacter sp. Py2] AAL15300 AT4g14210/dl3145c [Arabidopsis thaliana] AAL38046 phytoene desaturase [Hordeum vulgare] AAL73986 hypothetical protein MMT-7 [Mus musculus] AAL80005 phytoene desaturase [Sandersonia aurantiaca] AAL91366 carotenoid isomerase [Lycopersicon esculentum] AAM45380 phytoene desaturase [Tagetes erecta] AAM48646 phytoene dehydrogenase [uncultured proteobacterium] AAM63349 putative zeta-carotene desaturase precursor [Arabidopsis thaliana] AAM94364 phytoene desaturase [Agromyces mediolanus] AAN75037 CrtD [Rhodospirillum rubrum] AAN85599 Phytoene Desaturase [Pantoea stewartii] AAO24235 phytoene desaturase [Crocus sativus] AAO46892 phytoene dehydrogenase [Blakeslea trispora] AAO46894 phytoene dehydrogenase [Blakeslea trispora] AAO53257 phytoene desaturase [Xanthophyllomyces dendrorhous] AAO53258 phytoene desaturase splice variant [Xanthophyllomyces dendrorhous] AAO64750 At5g49550/K6M13_10 [Arabidopsis thaliana] AAO93135 phytoene dehydrogenase; CrtI [Rubrivivax gelatinosus] AAP59036 CrtD [Thiocapsa roseopersicina] AAP79175 phytoene dehydrogenase [Bigelowiella natans] AAQ04224 zeta-carotene desaturase ZDS1 [Malus × domestica] AAQ04225 zeta-carotene desaturase ZDS2 [Malus × domestica] AAQ65246 phytoene desaturase [Methylobacterium extorquens] AAQ65246 phytoene desaturase [Methylobacterium extorquens] AAQ88931 WLPL439 [Homo sapiens] AAR37797 methoxyneurosporene dehydrogenase [uncultured bacterium 442] AAR37802 phytoene dehydrogenase [uncultured bacterium 442] AAR37850 methoxyneurosporene dehydrogenase [uncultured bacterium 443] AAR37855 phytoene dehydrogenase [uncultured bacterium 443] AAR86105 phytoene desaturase [Momordica charantia var. abbreviata] AAR98491 phytoene desaturase [Bradyrhizobium sp. ORS278] AAR98494 phytoene dehydrogenase [Bradyrhizobium sp. ORS278] AAR98733 phytoene desaturase [Lilium longiflorum] AAS17750 phytoene desaturase [Solanum tuberosum] AAT01639 phytoene desaturase [Petunia × hybrida] AAT35222 fusion of carotene synthesis proteins [synthetic construct] AAT74579 PDS [Citrus sinensis] AAT74580 ZDS [Citrus sinensis] AAT76050 zeta-carotene desaturase [Citrus clementina] AAT76434 phytoene desaturase [Hydrilla verticillata] AAT90316 putative methoxyneurosporene dehydrogenase [uncultured proteobacterium eBACred25D05] AAU34019 retina and RPE/choroid [Macaca fascicularis] AAW23161 phytoene dehydrogenase [Rhodococcus erythropolis] AB2035 phytoene desaturase [imported] - Nostoc sp. (strain PCC 7120) AB2064 hypothetical protein alr2064 [imported] - Nostoc sp. (strain PCC 7120) AC2446 hypothetical protein all5123 [imported] - Nostoc sp. (strain PCC 7120) AF1557 phytoene dehydrogenase homolog lin0999 [imported] - Listeria innocua (strain Clip11262) AF2029 hypothetical protein alr1788 [imported] - Nostoc sp. (strain PCC 7120) AG2103 zeta-carotene desaturase [imported] - Nostoc sp. (strain PCC 7120) AG2509 zeta-carotene desaturase [imported] - Nostoc sp. (strain PCC 7120) plasmid pCC7120alpha AH1199 phytoene dehydrogenase homolog lmo1000 [imported] - Listeria monocytogenes (strain EGD-e) AI2185 hypothetical protein all3040 [imported] - Nostoc sp. (strain PCC 7120) AI2273 hypothetical protein all3744 [imported] - Nostoc sp. (strain PCC 7120) B55548 crtN protein - Staphylococcus aureus B84327 phytoene dehydrogenase [imported] - Halobacterium sp. NRC-1 B90061 squalene synthase [imported] - Staphylococcus aureus (strain N315) BAA14127 crtI [Pantoea ananatis] BAA20276 phytoene desaturase [Erythrobacter longus] BAA76534 Phytoene disaturase [Acidiphilium rubrum] BAB10768 phytoene dehydrogenase-like [Arabidopsis thaliana] BAB50520 phytoene dehydrogenase [Mesorhizobium loti MAFF303099] BAB51896 mlr5446 [Mesorhizobium loti MAFF303099] BAB68552 zeta-carotene desaturase [Citrus unshiu] BAB79603 crtI [Pantoea agglomerans pv. milletiae] BAB82461 phytoene desaturase [Gentiana lutea] BAB82462 phytoene desaturase [Gentiana lutea] BAB98016 Phytoene dehydrogenase and related proteins [Corynebacterium glutamicum ATCC 13032] BAC75676 putative phytoene desaturase [Gordonia sp. TM414] BAC77668 phytoene desaturase [marine bacterium P99-3] BAC77671 gamma-carotene desaturase [marine bacterium P99-3] BAD07279 phytoene desaturase [Citrus sinensis] BAD07280 zeta-carotene desaturase [Citrus sinensis] BAD07287 phytoene desaturase [Citrus limon] BAD07288 zeta-carotene desaturase [Citrus limon] CAA52098 squalene synthase [Staphylococcus aureus] CAA60479 Phytoene desaturase [Haematococcus pluvialis] CAA66626 unnamed protein product [Staphylococcus aureus] CAB38739 phytoene dehydrogenase [Rhodobacter sphaeroides] CAB38743 methoxyneurosporene dehydrogenase [Rhodobacter sphaeroides] CAB40843 phytoene dehydrogenase [Mucor circinelloides f. lusitanicus] CAB56041 zeta-carotene desaturase [Nostoc sp. PCC 7120] CAB56062 phytoene desaturase [Paracoccus marcusii] CAB59726 phytoene desaturase [Lycopersicon esculentum] CAB65434 zeta carotene desaturase [Synechococcus leopoliensis] CAB94794 phytoene dehydrogenase [Mycobacterium aurum] CAC85667 zeta-carotene desaturase [Citrus sinensis] CAD19989 phytoene dehydrogenase [Gibberella fujikuroi] CAD27442 putative zeta-carotene desaturase [Helianthus annuus] CAD55814 putative zeta-carotene desaturase [Helianthus annuus] CAE00192 phytoene desaturase [Nicotiana benthamiana] CAE83576 phytoene desaturase [Nicotiana tabacum] CAF19330 PHYTOENE DEHYDROGENASE (DESATURASE) [Corynebacterium glutamicum ATCC 13032] CAF21094 PHYTOENE DEHYDROGENASE (DESATURASE) (N-terminal fragment) [Corynebacterium glutamicum ATCC 13032] CAF21337 phytoene desaturase [Pisum sativum] CAH91165 hypothetical protein [Pongo pygmaeus] E90061 hypothetical protein SA2351 [imported] - Staphylococcus aureus (strain N315) EAA90383 unknown [environmental sequence] EAA98598 unknown [environmental sequence] EAB09790 unknown [environmental sequence] EAB14136 unknown [environmental sequence] EAB18725 unknown [environmental sequence] EAB29729 unknown [environmental sequence] EAB30992 unknown [environmental sequence] EAB41377 unknown [environmental sequence] EAB54727 unknown [environmental sequence] EAB76679 unknown [environmental sequence] EAB87028 unknown [environmental sequence] EAB92587 unknown [environmental sequence] EAB94459 unknown [environmental sequence] EAB96864 unknown [environmental sequence] EAC01884 unknown [environmental sequence] EAC38895 unknown [environmental sequence] EAC60360 unknown [environmental sequence] EAD05874 unknown [environmental sequence] EAD05999 unknown [environmental sequence] EAD20520 unknown [environmental sequence] EAE06978 unknown [environmental sequence] EAE70773 unknown [environmental sequence] EAF04985 unknown [environmental sequence] EAF51354 unknown [environmental sequence] EAF62819 unknown [environmental sequence] EAF75453 unknown [environmental sequence] EAG09111 unknown [environmental sequence] EAG19412 unknown [environmental sequence] EAG23070 unknown [environmental sequence] EAG25053 unknown [environmental sequence] EAG25054 unknown [environmental sequence] EAG29279 unknown [environmental sequence] EAG39845 unknown [environmental sequence] EAG56100 unknown [environmental sequence] EAG63013 unknown [environmental sequence] EAG68633 unknown [environmental sequence] EAG71574 unknown [environmental sequence] EAG89835 unknown [environmental sequence] EAH04928 unknown [environmental sequence] EAH04936 unknown [environmental sequence] EAH08586 unknown [environmental sequence] EAH22597 unknown [environmental sequence] EAH22853 unknown [environmental sequence] EAH31648 unknown [environmental sequence] EAH55579 unknown [environmental sequence] EAH68071 unknown [environmental sequence] EAH73302 unknown [environmental sequence] EAH79041 unknown [environmental sequence] EAH86965 unknown [environmental sequence] EAH97108 unknown [environmental sequence] EAH99977 unknown [environmental sequence] EAI01660 unknown [environmental sequence] EAI03576 unknown [environmental sequence] EAI06784 unknown [environmental sequence] EAI11087 unknown [environmental sequence] EAI15261 unknown [environmental sequence] EAI15547 unknown [environmental sequence] EAI17521 unknown [environmental sequence] EAI21398 unknown [environmental sequence] EAI29728 unknown [environmental sequence] EAI38468 unknown [environmental sequence] EAI43591 unknown [environmental sequence] EAI51589 unknown [environmental sequence] EAI58453 unknown [environmental sequence] EAI72974 unknown [environmental sequence] EAI77885 unknown [environmental sequence] EAI78272 unknown [environmental sequence] EAI80262 unknown [environmental sequence] EAI83937 unknown [environmental sequence] EAI86664 unknown [environmental sequence] EAJ00517 unknown [environmental sequence] EAJ05570 unknown [environmental sequence] EAJ08238 unknown [environmental sequence] EAJ15524 unknown [environmental sequence] EAJ18144 unknown [environmental sequence] EAJ20649 unknown [environmental sequence] EAJ21683 unknown [environmental sequence] EAJ24413 unknown [environmental sequence] EAJ28774 unknown [environmental sequence] EAJ30522 unknown [environmental sequence] EAJ35157 unknown [environmental sequence] EAJ37407 unknown [environmental sequence] EAJ39929 unknown [environmental sequence] EAJ54356 unknown [environmental sequence] EAJ54959 unknown [environmental sequence] EAJ56207 unknown [environmental sequence] EAJ58447 unknown [environmental sequence] EAJ59958 unknown [environmental sequence] EAJ63347 unknown [environmental sequence] EAJ66054 unknown [environmental sequence] EAJ67637 unknown [environmental sequence] EAJ69812 unknown [environmental sequence] EAJ74441 unknown [environmental sequence] EAJ76472 unknown [environmental sequence] EAJ76473 unknown [environmental sequence] EAJ80355 unknown [environmental sequence] EAJ80839 unknown [environmental sequence] EAJ81408 unknown [environmental sequence] EAJ86174 unknown [environmental sequence] EAJ87600 unknown [environmental sequence] EAJ88203 unknown [environmental sequence] EAJ88682 unknown [environmental sequence] EAJ92341 unknown [environmental sequence] EAJ94774 unknown [environmental sequence] EAJ97555 unknown [environmental sequence] EAJ97958 unknown [environmental sequence] EAK07654 unknown [environmental sequence] EAK08513 unknown [environmental sequence] EAK08529 unknown [environmental sequence] EAK10609 unknown [environmental sequence] EAK10614 unknown [environmental sequence] EAK12902 unknown [environmental sequence] EAK13034 unknown [environmental sequence] EAK15092 unknown [environmental sequence] EAK22483 unknown [environmental sequence] EAK23222 unknown [environmental sequence] EAK24187 unknown [environmental sequence] EAK24674 unknown [environmental sequence] EAK28785 unknown [environmental sequence] EAK34731 unknown [environmental sequence] EAK34742 unknown [environmental sequence] EAK36883 unknown [environmental sequence] EAK37522 unknown [environmental sequence] EAK42705 unknown [environmental sequence] EAK43213 unknown [environmental sequence] EAK52580 unknown [environmental sequence] EAK53452 unknown [environmental sequence] EAK58759 unknown [environmental sequence] EAK62665 unknown [environmental sequence] EAK63558 unknown [environmental sequence] F84187 phytoene dehydrogenase [imported] - Halobacterium sp. NRC-1 F90272 phytoene dehydrogenase related protein [imported] - Sulfolobus solfataricus G87635 phytoene dehydrogenase-related protein [imported] - Caulobacter crescentus G90413 phytoene dehydrogenase related protein [imported] - Sulfolobus solfataricus H83880 hypothetical protein BH1848 [imported] - Bacillus halodurans (strain C-125) H84320 phytoene dehydrogenase [imported] - Halobacterium sp. NRC-1 JC7723 phytoene desaturase (EC 1.14.99.—) 1 - citrus NP_060220 all-trans-13,14-dihydroretinol saturase [Homo sapiens] NP_080435 all-trans-13,14-dihydroretinol saturase [Mus musculus] NP_193157 phytoene dehydrogenase, chloroplast/phytoene desaturase (PDS) [Arabidopsis thaliana] NP_214383 protoporphyrinogen oxidase [Aquifex aeolicus VF5] NP_276913 phytoene dehydrogenase [Methanothermobacter thermautotrophicus str. Delta H] NP_293819 phytoene dehydrogenase, putative [Deinococcus radiodurans R1] NP_294534 dehydrogenase, putative [Deinococcus radiodurans R1] NP_294585 phytoene dehydrogenase [Deinococcus radiodurans R1] NP_295972 methoxyneurosporene dehydrogenase [Deinococcus radiodurans R1] NP_338490 oxidoreductase, putative [Mycobacterium tuberculosis CDC1551] NP_376437 hypothetical protein ST0549 [Sulfolobus tokodaii str. 7] NP_377056 hypothetical protein ST1130 [Sulfolobus tokodaii str. 7] NP_388895 protoporphyrinogen IX and coproporphyrinogen III oxidase [Bacillus subtilis subsp. subtilis str. 168] NP_441167 phytoene desaturase [Synechocystis sp. PCC 6803] NP_441254 hypothetical protein sll0254 [Synechocystis sp. PCC 6803] NP_442491 b-carotene ketolase [Synechocystis sp. PCC 6803] NP_442727 hypothetical protein sll0033 [Synechocystis sp. PCC 6803] NP_562475 probable diapophytoene dehydrogenase [Clostridium perfringens str. 13] NP_568712 amine oxidase-related [Arabidopsis thaliana] NP_601630 hypothetical protein NCgl2346 [Corynebacterium glutamicum ATCC 13032] NP_601630 hypothetical protein NCgl2346 [Corynebacterium glutamicum ATCC 13032] NP_616426 phytoene dehydrogenase family protein [Methanosarcina acetivorans C2A] NP_624522 putative phytoene dehydrogenase (phytoene desaturase) (putative secreted protein) [Streptomyces coelicolor A3(2)] NP_626360 putative carotenoid dehydrogenase (putative secreted protein) [Streptomyces coelicolor A3(2)] NP_630834 putative phytoene dehydrogenase [Streptomyces coelicolor A3(2)] NP_643053 phytoene dehydrogenase [Xanthomonas axonopodis pv. citri str. 306] NP_647302 hypothetical protein MW2485 [Staphylococcus aureus subsp. aureus MW2] NP_659552 all-trans-13,14-dihydroretinol saturase [Rattus norvegicus] NP_661086 lycopene cyclase, putative [Chlorobium tepidum TLS] NP_661546 carotenoid isomerase, putative [Chlorobium tepidum TLS] NP_661701 phytoene desaturase [Chlorobium tepidum TLS] NP_662300 zeta-carotene desaturase [Chlorobium tepidum TLS] NP_681023 putative phytoene dehydrogenase [Thermosynechococcus elongatus BP-1] NP_681127 zeta-carotene desaturase [Thermosynechococcus elongatus BP-1] NP_682351 phytoene dehydrogenase/phytoene desaturase [Thermosynechococcus elongatus BP-1] NP_693380 phytoene dehydrogenase [Oceanobacillus iheyensis HTE831] NP_693382 phytoene dehydrogenase [Oceanobacillus iheyensis HTE831] NP_737250 phytoene desaturase [Corynebacterium efficiens YS-314] NP_763380 hypothetical protein VV21505 [Vibrio vulnificus CMCP6] NP_786524 squalene synthase [Lactobacillus plantarum WCFS1] NP_822198 phytoene desaturase [Streptomyces avermitilis MA-4680] NP_822828 squalene/phytoene dehydrogenase [Streptomyces avermitilis MA-4680] NP_827278 putative phytoene dehydrogenase [Streptomyces avermitilis MA-4680] NP_851528 putative phytoene dehydrogenase [Streptomyces rochei] NP_857496 PUTATIVE DEHYDROGENASE [Mycobacterium bovis AF2122/97] NP_868798 probable phytoene dehydrogenase [Rhodopirellula baltica SH 1] NP_869339 phytoene dehydrogenase [Rhodopirellula baltica SH 1] NP_870237 phytoene dehydrogenase [Rhodopirellula baltica SH 1] NP_874530 Zeta-carotene desaturase [Prochlorococcus marinus subsp. marinus str. CCMP1375] NP_874561 Phytoene dehydrogenase, phytoene desaturase [Prochlorococcus marinus subsp. marinus str. CCMP1375] NP_874977 Phytoene dehydrogenase/carotenoid isomerase [Prochlorococcus marinus subsp. marinus str. CCMP1375] NP_892236 zeta-carotene desaturase [Prochlorococcus marinus subsp. pastoris str. CCMP1986] NP_892265 phytoene desaturase [Prochlorococcus marinus subsp. pastoris str. CCMP1986] NP_892458 Bacterial-type phytoene dehydrogenase [Prochlorococcus marinus subsp. pastoris str. CCMP1986] NP_893232 putative carotenoid isomerase [Prochlorococcus marinus subsp. pastoris str. CCMP1986] NP_894882 putative carotenoid isomerase [Prochlorococcus marinus str. MIT 9313] NP_895385 NAD binding site [Prochlorococcus marinus str. MIT 9313] NP_895793 zeta-carotene desaturase [Prochlorococcus marinus str. MIT 9313] NP_895829 phytoene desaturase [Prochlorococcus marinus str. MIT 9313] NP_896854 hypothetical protein SYNW0761 [Synechococcus sp. WH 8102] NP_896994 Carotenoid isomerase [Synechococcus sp. WH 8102] NP_898304 zeta-carotene desaturase [Synechococcus sp. WH 8102] NP_898346 phytoene desaturase [Synechococcus sp. WH 8102] NP_902647 hypothetical protein CV2977 [Chromobacterium violaceum ATCC 12472] NP_923340 beta-carotene ketolase [Gloeobacter violaceus PCC 7421] NP_923639 hypothetical protein gll0693 [Gloeobacter violaceus PCC 7421] NP_923813 phytoene dehydrogenase [Gloeobacter violaceus PCC 7421] NP_925079 hypothetical protein gvip293 [Gloeobacter violaceus PCC 7421] NP_931515 phytoene dehydrogenase (phytoene desaturase) [Photorhabdus luminescens subsp. laumondii TTO1] NP_936379 hypothetical protein VVA0323 [Vibrio vulnificus YJ016] NP_940208 phytoene dehydrogenase related enzyme [Corynebacterium diphtheriae NCTC 13129] NP_945754 NAD binding site:Amine oxidase [Rhodopseudomonas palustris CGA009] NP_946860 phytoene dehydrogenase CrtI [Rhodopseudomonas palustris CGA009] NP_946866 methoxyneurosporene dehydrogenase [Rhodopseudomonas palustris CGA009] NP_948851 phytoene dehydrogenase-related protein [Rhodopseudomonas palustris CGA009] NP_962004 hypothetical protein MAP3070 [Mycobacterium avium subsp. paratuberculosis str. k10] NP_968600 Phytoene dehydrogenase [Bdellovibrio bacteriovorus HD100] NP_974222 zeta-carotene desaturase (ZDS1)/carotene 7,8-desaturase [Arabidopsis thaliana] NP_974545 phytoene dehydrogenase, chloroplast/phytoene desaturase (PDS) [Arabidopsis thaliana] O49901 Zeta-carotene desaturase, chloroplast precursor (Carotene 7,8-desaturase) P17059 Methoxyneurosporene dehydrogenase P54971 Phytoene dehydrogenase (Phytoene desaturase) P54978 Phytoene dehydrogenase (Phytoene desaturase) P54979 Phytoene dehydrogenase (Phytoene desaturase) P54981 Phytoene dehydrogenase (Phytoene desaturase) P54982 Phytoene dehydrogenase (Phytoene desaturase) P74306 Zeta-carotene desaturase (Carotene 7,8-desaturase) Q01671 Methoxyneurosporene dehydrogenase Q02861 Phytoene dehydrogenase (Phytoene desaturase) Q38893 Zeta-carotene desaturase, chloroplast precursor (Carotene 7,8-desaturase) Q40406 Phytoene dehydrogenase, chloroplast precursor (Phytoene desaturase) Q9FV46 Zeta-carotene desaturase, chloroplast precursor (Carotene 7,8-desaturase) Q9SE20 Zeta-carotene desaturase, chloroplast precursor (Carotene 7,8-desaturase) Q9SMJ3 Zeta-carotene desaturase, chloroplast precursor (Carotene 7,8-desaturase) Q9ZTN9 Phytoene dehydrogenase, chloroplast precursor (Phytoene desaturase) Q9ZTP4 Zeta-carotene desaturase, chloroplast precursor (Carotene 7,8-desaturase) S29314 phytoene dehydrogenase (EC 1.3.—.—) - pepper S32171 hydroxyneurosporene dehydrogenase (EC 1.3.—.—) - Myxococcus xanthus S49624 methoxyneurosporene dehydrogenase (EC 1.3.—.—) CrtD - Rhodobacter sphaeroides S52586 phytoene dehydrogenase (EC 1.3.—.—) - Erwinia herbicola S65060 phytoene desaturase precursor - maize T10701 probable phytoene dehydrogenase (EC 1.3.—.—) - green alga (Dunaliella bardawil) T31463 probable diapophytoene dehydrogenase crtN - Heliobacillus mobilis T46822 phytoene desaturase (EC 1.3.—.—) [validated] - Xanthophyllomyces dendrorhous T48646 phytoene dehydrogenase (EC 1.—.—.—) [validated] - Cercospora nicotianae T50745 phytoene dehydrogenase (EC 1.3.—.—) [imported] - Rhodobacter sphaeroides T50749 methoxyneurosporene dehydrogenase (EC 1.3.—.—) crtD [imported] - Rhodobacter sphaeroides T50893 methoxyneurosporene dehydrogenase [imported] - Rubrivivax gelatinosus T50910 phytoene dehydrogenase [imported] - Rubrivivax gelatinosus T51119 phytoene desaturase crtI [imported] - Brevibacterium linens T51123 betacarotene desaturase [imported] - Brevibacterium linens XP_324732 PHYTOENE DEHYDROGENASE (PHYTOENE DESATURASE) (ALBINO-1 PROTEIN) [Neurospora crassa] XP_383241 hypothetical protein FG03065.1 [Gibberella zeae PH-1] XP_401825 hypothetical protein UM04210.1 [Ustilago maydis 521] XP_470568 Putative phytoene dehydrogenase precursor [Oryza sativa] XP_473486 OSJNBa0084K11.8 [Oryza sativa (japonica cultivar-group)] XP_477063 putative zeta-carotene desaturase precursor [Oryza sativa (japonica cultivar-group)] XP_525801 PREDICTED: hypothetical protein XP_525801 [Pan troglodytes] XP_540198 PREDICTED: hypothetical protein XP_540198 [Canis familiaris] YP_006049 phytoene dehydrogenase [Thermus thermophilus HB27] YP_013621 phytoene dehydrogenase, putative [Listeria monocytogenes str. 4b F2365] YP_024310 phytoene dehydrogenase [Picrophilus torridus DSM 9790] YP_041986 squalene synthase [Staphylococcus aureus subsp. aureus MRSA252] YP_041988 putative phytoene dehydrogenase related protein [Staphylococcus aureus subsp. aureus MRSA252] YP_044561 squalene synthase [Staphylococcus aureus subsp. aureus MSSA476] YP_044564 putative phytoene dehydrogenase related protein [Staphylococcus aureus subsp. aureus MSSA476] YP_062471 phytoene dehydrogenase [Leifsonia xyli subsp. xyli str. CTCB07] YP_117947 putative phytoene desaturase [Nocardia farcinica IFM 10152] YP_120612 putative phytoene desaturase [Nocardia farcinica IFM 10152] YP_135077 phytoene dehydrogenase [Haloarcula marismortui ATCC 43049] YP_136483 phytoene dehydrogenase [Haloarcula marismortui ATCC 43049] YP_145331 probable phytoene dehydrogenase [Thermus thermophilus HB8] YP_145348 phytoene dehydrogenase [Thermus thermophilus HB8] YP_171014 carotene isomerase [Synechococcus elongatus PCC 6301] YP_172823 phytoene dehydrogenase [Synechococcus elongatus PCC 6301] YP_173078 carotene isomerase [Synechococcus elongatus PCC 6301] YP_173207 zeta-carotene desaturase [Synechococcus elongatus PCC 6301] YP_184572 predicted oxidoreductase [Thermococcus kodakaraensis] YP_187368 dehydrosqualene desaturase [Staphylococcus aureus subsp. aureus COL] YP_187371 phytoene dehydrogenase [Staphylococcus aureus subsp. aureus COL] YP_187371 phytoene dehydrogenase [Staphylococcus aureus subsp. aureus COL] YP_187371 phytoene dehydrogenase [Staphylococcus aureus subsp. aureus COL] ZP_000490 COG1233: Phytoene dehydrogenase and related proteins [Magnetospirillum magnetotacticum MS-1] ZP_000509 COG1233: Phytoene dehydrogenase and related proteins [Magnetospirillum magnetotacticum MS-1] ZP_000518 COG1233: Phytoene dehydrogenase and related proteins [Magnetospirillum magnetotacticum MS-1] ZP_000566 COG0562: UDP-galactopyranose mutase [Magnetospirillum magnetotacticum MS- 1] ZP_000627 COG1233: Phytoene dehydrogenase and related proteins [Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293] ZP_000627 COG1233: Phytoene dehydrogenase and related proteins [Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293] ZP_001073 COG1233: Phytoene dehydrogenase and related proteins [Nostoc punctiforme PCC 73102] ZP_001081 COG1233: Phytoene dehydrogenase and related proteins [Nostoc punctiforme PCC 73102] ZP_001091 COG3349: Uncharacterized conserved protein [Nostoc punctiforme PCC 73102] ZP_001116 COG1233: Phytoene dehydrogenase and related proteins [Nostoc punctiforme PCC 73102] ZP_001117 COG3349: Uncharacterized conserved protein [Nostoc punctiforme PCC 73102] ZP_001119 COG3349: Uncharacterized conserved protein [Nostoc punctiforme PCC 73102] ZP_001124 COG1233: Phytoene dehydrogenase and related proteins [Nostoc punctiforme PCC 73102] ZP_001510 COG1233: Phytoene dehydrogenase and related proteins [Dechloromonas aromatica RCB] ZP_001591 COG3349: Uncharacterized conserved protein [Anabaena variabilis ATCC 29413] ZP_001593 COG1233: Phytoene dehydrogenase and related proteins [Anabaena variabilis ATCC 29413] ZP_001602 COG3349: Uncharacterized conserved protein [Anabaena variabilis ATCC 29413] ZP_001614 COG3349: Uncharacterized conserved protein [Anabaena variabilis ATCC 29413] ZP_001645 COG3349: Uncharacterized conserved protein [Synechococcus elongatus PCC 7942] ZP_001650 COG3349: Uncharacterized conserved protein [Synechococcus elongatus PCC 7942] ZP_001722 COG1233: Phytoene dehydrogenase and related proteins [Methylobacillus flagellatus KT] ZP_001746 COG3349: Uncharacterized conserved protein [Crocosphaera watsonii WH 8501] ZP_001752 COG3349: Uncharacterized conserved protein [Crocosphaera watsonii WH 8501] ZP_001770 COG1232: Protoporphyrinogen oxidase [Crocosphaera watsonii WH 8501] ZP_001777 COG1233: Phytoene dehydrogenase and related proteins [Crocosphaera watsonii WH 8501] ZP_001787 COG3349: Uncharacterized conserved protein [Crocosphaera watsonii WH 8501] ZP_001837 COG1233: Phytoene dehydrogenase and related proteins [Exiguobacterium sp. 255- 15] ZP_001867 COG1233: Phytoene dehydrogenase and related proteins [Rubrobacter xylanophilus DSM 9941] ZP_002073 COG1233: Phytoene dehydrogenase and related proteins [Rhodobacter sphaeroides 2.4.1] ZP_002077 COG1233: Phytoene dehydrogenase and related proteins [Rhodobacter sphaeroides 2.4.1] ZP_002339 phytoene dehydrogenase, putative [Listeria monocytogenes str. 1/2a F6854] ZP_002680 COG1233: Phytoene dehydrogenase and related proteins [Rhodospirillum rubrum] ZP_002705 COG1233: Phytoene dehydrogenase and related proteins [Rhodospirillum rubrum] ZP_002771 COG1232: Protoporphyrinogen oxidase [Burkholderia fungorum LB400] ZP_002892 hypothetical protein Mmc102002317 [Magnetococcus sp. MC-1] ZP_002916 COG3349: Uncharacterized conserved protein [Thermobifida fusca] ZP_002963 COG1233: Phytoene dehydrogenase and related proteins [Methanosarcina barkeri str. fusaro] ZP_003022 COG1233: Phytoene dehydrogenase and related proteins [Novosphingobium aromaticivorans DSM 12444] ZP_003036 COG1233: Phytoene dehydrogenase and related proteins [Novosphingobium aromaticivorans DSM 12444] ZP_003107 COG1233: Phytoene dehydrogenase and related proteins [Cytophaga hutchinsonii] ZP_003202 COG1233: Phytoene dehydrogenase and related proteins [Oenococcus oeni PSU-1] ZP_003258 COG1233: Phytoene dehydrogenase and related proteins [Trichodesmium erythraeum IMS101] ZP_003268 COG3349: Uncharacterized conserved protein [Trichodesmium erythraeum IMS101] ZP_003269 COG3349: Uncharacterized conserved protein [Trichodesmium erythraeum IMS101] ZP_003276 COG1233: Phytoene dehydrogenase and related proteins [Trichodesmium erythraeum IMS101] ZP_003283 COG1233: Phytoene dehydrogenase and related proteins [Trichodesmium erythraeum IMS101] ZP_003557 COG1233: Phytoene dehydrogenase and related proteins [Exiguobacterium sp. 255- 15] ZP_003559 COG3349: Uncharacterized conserved protein [Chloroflexus aurantiacus] ZP_003565 COG1233: Phytoene dehydrogenase and related proteins [Chloroflexus aurantiacus] ZP_003577 COG1233: Phytoene dehydrogenase and related proteins [Chloroflexus aurantiacus] ZP_003593 COG1233: Phytoene dehydrogenase and related proteins [Chloroflexus aurantiacus] ZP_003595 COG1233: Phytoene dehydrogenase and related proteins [Chloroflexus aurantiacus] ZP_003685 carotenoid isomerase, putative [Campylobacter lari RM2100]

TABLE 17b Examples of phytoene dehydrogenase polypeptides. Row Accession GI Protein Description 1 1613414B 227039 crtI gene 2 CAA36533 45998 unnamed protein product [Rhodobacter capsulatus] 3 ABP69925 145555312 Phytoene dehydrogenase-related protein [Rhodobacter sphaeroides ATCC 17025] 4 ABG29855 109453650 phytoene dehydrogenase [Roseobacter denitrificans OCh 114] 5 YP_001 . . . 126462676 Phytoene dehydrogenase-related protein [Rhodobacter sphaeroides ATCC 17029] 6 CAB38739 4490589 phytoene dehydrogenase [Rhodobacter sphaeroides] 7 AAF24289 6690721 CrtI [Rhodobacter sphaeroides] 8 ZP_019 . . . 149914257 Phytoene dehydrogenase [Roseobacter sp. AzwK-3b] 9 ZP_018 . . . 149203764 Phytoene dehydrogenase [Roseovarius sp. TM1035] 10 ZP_017 . . . 126734359 Phytoene dehydrogenase [Roseobacter sp. CCS2] 11 ZP_015 . . . 118735466 Phytoene dehydrogenase-related protein [Dinoroseobacter shibae DFL 12] 12 EBA75398 134340305 hypothetical protein GOS_348183 [marine metagenome] 13 ZP_010 . . . 85703533 Phytoene dehydrogenase [Roseovarius sp. 217] 14 ZP_010 . . . 84514966 Phytoene dehydrogenase [Loktanella vestfoldensis SKA53] 15 EDJ26541 144155062 hypothetical protein GOS_1726264 [marine metagenome] 16 AAM48646 21328640 phytoene dehydrogenase [uncultured proteobacterium] 17 EBF29285 135068871 hypothetical protein GOS_9621672 [marine metagenome] 18 EDH88525 143962357 hypothetical protein GOS_524801 [marine metagenome] 19 ABD53060 88862183 phytoene dehydrogenase [Jannaschia sp. CCS1] 20 ECX23858 142320992 hypothetical protein GOS_2574166 [marine metagenome] 21 EDD83467 143241615 hypothetical protein GOS_1236491 [marine metagenome] 22 ECW60413 142235101 hypothetical protein GOS_2690740 [marine metagenome] 23 EDB24382 142880994 hypothetical protein GOS_1857800 [marine metagenome] 24 EDC65629 143078396 hypothetical protein GOS_1437717 [marine metagenome] 25 EDC66763 143079940 hypothetical protein GOS_1435710 [marine metagenome] 26 EBF73535 135137918 hypothetical protein GOS_9549173 [marine metagenome] 27 EDB61897 142933299 hypothetical protein GOS_1624464 [marine metagenome] 28 EDH58399 143920581 hypothetical protein GOS_579274 [marine metagenome] 29 ECV46406 142084909 hypothetical protein GOS_2893885 [marine metagenome] 30 EBG03643 135184853 hypothetical protein GOS_9500102 [marine metagenome] 31 ECW11301 142170256 hypothetical protein GOS_2778421 [marine metagenome] 32 EDG30167 143690631 hypothetical protein GOS_805431 [marine metagenome] 33 EBF89344 135162533 hypothetical protein GOS_9523433 [marine metagenome] 34 EDG14791 143664156 hypothetical protein GOS_832322 [marine metagenome] 35 ECW44302 142213461 hypothetical protein GOS_2720548 [marine metagenome] 36 EDC28179 143027347 hypothetical protein GOS_1503868 [marine metagenome] 37 EBF91518 135165888 hypothetical protein GOS_9519783 [marine metagenome] 38 EBG06098 135188689 hypothetical protein GOS_9496261 [marine metagenome] 39 YP_001 . . . 121998398 Phytoene dehydrogenase-related protein [Halorhodospira halophila SL1] 40 ABC21298 83574747 Phytoene desaturase [Rhodospirillum rubrum ATCC 11170] 41 EDD45555 143188703 hypothetical protein GOS_1301547 [marine metagenome] 42 EBF89879 135163364 hypothetical protein GOS_9522488 [marine metagenome] 43 EDA94958 142836512 hypothetical protein GOS_1907603 [marine metagenome] 44 EBE40240 134934911 hypothetical protein GOS_9769793 [marine metagenome] 45 ABQ38355 146409849 Phytoene dehydrogenase (Phytoene desaturase) [Bradyrhizobium sp. BTAi1] 46 ECV53668 142093711 hypothetical protein GOS_2880588 [marine metagenome] 47 EDJ05722 144126667 hypothetical protein GOS_1762374 [marine metagenome] 48 EDJ52003 144189327 hypothetical protein GOS_1681048 [marine metagenome] 49 CAL75489 146191484 Phytoene dehydrogenase (Phytoene desaturase) [Bradyrhizobium sp. ORS278] 50 ZP_011 . . . 88706670 phytoene dehydrogenase [gamma proteobacterium KT 71] 51 ABD86823 90104786 amine oxidase [Rhodopseudomonas palustris BisB18] 52 ZP_019 . . . 149922641 amine oxidase [Plesiocystis pacifica SIR-1] 53 ECW14126 142173943 hypothetical protein GOS_2773516 [marine metagenome] 54 ABJ05267 115517283 amine oxidase [Rhodopseudomonas palustris BisA53] 55 ZP_020 . . . 156449192 Zeta-phytoene desaturase [Methylobacterium chloromethanicum CM4] 56 ZP_020 . . . 153900167 FAD-dependent pyridine nucleotide-disulphide oxidoreductase [Methylobacterium extorquens PA1] 57 EDO75157 157409160 Zeta-phytoene desaturase [Methylobacterium populi BJ001] 58 EBF26787 135064945 hypothetical protein GOS_9625603 [marine metagenome] 59 EBM47492 136188235 hypothetical protein GOS_8404015 [marine metagenome] 60 BAA94063 7416814 phytoene dehydrogenase [Rubrivivax gelatinosus] 61 AAO93135 29893506 phytoene dehydrogenase; CrtI [Rubrivivax gelatinosus] 62 ABE40987 91684685 amine oxidase [Rhodopseudomonas palustris BisB5] 63 ABC44467 83756354 phytoene dehydrogenase [Salinibacter ruber DSM 13855] 64 NP_946860 39934584 phytoene dehydrogenase CrtI [Rhodopseudomonas palustris CGA009] 65 EDK95498 148522573 Phytoene dehydrogenase-related protein [Methylobacterium sp. 4- 46] 66 AAX48189 61653236 phytoene dehydrogenase [uncultured proteobacterium DelRiverFos06H03] 67 ABD08704 86574147 Amine oxidase [Rhodopseudomonas palustris HaA2] 68 ECV97125 142151708 hypothetical protein GOS_2803067 [marine metagenome] 69 EBF91320 135165576 hypothetical protein GOS_9520100 [marine metagenome] 70 EDG74671 143780072 hypothetical protein GOS_728533 [marine metagenome] 71 AAR37802 40062931 phytoene dehydrogenase [uncultured bacterium 442] 72 ZP_016 . . . 119504131 amine oxidase [marine gamma proteobacterium HTCC2080] 73 EBY63975 138260458 hypothetical protein GOS_5033312 [marine metagenome] 74 ECW52975 142225047 hypothetical protein GOS_2704529 [marine metagenome] 75 EDB07926 142858428 hypothetical protein GOS_1885673 [marine metagenome] 76 ECX08477 142300457 hypothetical protein GOS_2602107 [marine metagenome] 77 EBE08763 134887517 hypothetical protein GOS_9823078 [marine metagenome] 78 EDG53739 143740094 hypothetical protein GOS_764800 [marine metagenome] 79 AAR37855 40062985 phytoene dehydrogenase [uncultured bacterium 443] 80 EBE55393 134957668 hypothetical protein GOS_9744377 [marine metagenome] 81 EBK49247 135888345 hypothetical protein GOS_8723699 [marine metagenome] 82 YP_001 . . . 154247504 Zeta-phytoene desaturase [Xanthobacter autotrophicus Py2] 83 EBJ38620 135714057 hypothetical protein GOS_8907038 [marine metagenome] 84 EBO29607 136458570 hypothetical protein GOS_8104065 [marine metagenome] 85 EDI63495 144067673 hypothetical protein GOS_399405 [marine metagenome] 86 ECH51213 139794606 hypothetical protein GOS_4963038 [marine metagenome] 87 ZP_014 . . . 113941052 amine oxidase [Herpetosiphon aurantiacus ATCC 23779] 88 P54978 85700409 Phytoene dehydrogenase (Phytoene desaturase) 89 AAY28420 63034218 phytoene desaturase [Paracoccus haeundaensis] 90 YP_001 . . . 148655097 Phytoene dehydrogenase-related protein [Roseiflexus sp. RS-1] 91 AAT35222 47531118 fusion of carotene synthesis proteins [synthetic construct] 92 CAB56062 5912295 phytoene desaturase [Paracoccus marcusii] 93 ZP_016 . . . 119484684 phytoene dehydrogenase CrtI [Lyngbya sp. PCC 8106] 94 YP_001 . . . 156932562 hypothetical protein ESA_00343 [Enterobacter sakazakii ATCC BAA-894] 95 CAL34121 112702901 phytoene dehydrogenase CRTI [Enterobacter sakazakii] 96 ZP_014 . . . 114705593 phytoene dehydrogenase (phytoene desaturase) [Fulvimarina pelagi HTCC2506] 97 ABU57678 156232895 Zeta-phytoene desaturase [Roseiflexus castenholzii DSM 13941] 98 EDJ24172 144151864 hypothetical protein GOS_1730347 [marine metagenome] 99 EBP04886 136575764 hypothetical protein GOS_7976019 [marine metagenome] 100 EDJ19941 144146057 hypothetical protein GOS_1737754 [marine metagenome] 101 EDE02809 143268695 hypothetical protein GOS_1202430 [marine metagenome] 102 EDA02972 142711842 hypothetical protein GOS_2076612 [marine metagenome] 103 AAC44850 1842244 phytoene desaturase 104 ECI48880 139936452 hypothetical protein GOS_4592564 [marine metagenome] 105 ECW29436 142194003 hypothetical protein GOS_2746710 [marine metagenome] 106 CAE79593 39575425 Phytoene dehydrogenase [Bdellovibrio bacteriovorus HD100] 107 EDJ16294 144141089 hypothetical protein GOS_1744028 [marine metagenome] 108 ABC50114 84043362 phytoene desaturase [Brevundimonas vesicularis] 109 CAE16714 36787607 phytoene dehydrogenase (phytoene desaturase) [Photorhabdus luminescens subsp. laumondii TTO1] 110 AAZ73142 72536076 phytoene desaturase [Enterobacteriaceae bacterium DC416] 111 ECC15254 138850366 hypothetical protein GOS_5005338 [marine metagenome] 112 EBF96527 135173721 hypothetical protein GOS_9511771 [marine metagenome] 113 AAZ73130 72536062 phytoene desaturase [Enterobacteriaceae bacterium DC260] 114 AAW72814 92430259 CrtI [Pantoea sp. C1B1Y] 115 EBF63722 135122709 hypothetical protein GOS_9565260 [marine metagenome] 116 EDJ33653 144164568 hypothetical protein GOS_1713484 [marine metagenome] 117 BAB79603 18143448 crtI [Pantoea agglomerans pv. milletiae] 118 AAZ73136 72536069 phytoene desaturase [Enterobacteriaceae bacterium DC404] 119 ABD73281 89475493 phytoene dehydrogenase [Pantoea agglomerans] 120 AAF78201 8650417 phytoene desaturase [Bradyrhizobium sp. ORS278] 121 ZP_010 . . . 84704558 phytoene dehydrogenase (phytoene desaturase) [Parvularcula bermudensis HTCC2503] 122 EBF30985 135071529 hypothetical protein GOS_9618780 [marine metagenome] 123 EDJ11107 144133947 hypothetical protein GOS_1752989 [marine metagenome] 124 YP_001 . . . 146284185 phytoene desaturase [Pseudomonas stutzeri A1501] 125 EDB39933 142902995 hypothetical protein GOS_1830750 [marine metagenome] 126 ZP_013 . . . 94496006 amine oxidase [Sphingomonas sp. SKA58] 127 P22871 117514 Phytoene dehydrogenase (Phytoene desaturase) 128 ECD12151 139058277 hypothetical protein GOS_4688757 [marine metagenome] 129 AAA64981 551855 phytoene dehydrogenase [Pantoea agglomerans] 130 ECS36705 141558903 hypothetical protein GOS_4990354 [marine metagenome] 131 ECJ26724 140055135 hypothetical protein GOS_5004115 [marine metagenome] 132 ECY06051 142435846 hypothetical protein GOS_2424849 [marine metagenome] 133 ZP_007 . . . 76261240 Amine oxidase: FAD dependent oxidoreductase [Chloroflexus aurantiacus J-10-fl] 134 BAD99408 67003497 phytoene desaturase [Brevundimonas sp. SD212] 135 ZP_015 . . . 118048084 amine oxidase [Chloroflexus aggregans DSM 9485] 136 EAS49801 90336053 phytoene dehydrogenase [Aurantimonas sp. SI85-9A1] 137 EDF88410 143616932 hypothetical protein GOS_877736 [marine metagenome] 138 AAN85599 27228293 Phytoene Desaturase [Pantoea stewartii] 139 EDI65706 144070706 hypothetical protein GOS_395761 [marine metagenome] 140 AAA21263 148397 phytoene dehydrogenase [Pantoea agglomerans] 141 P21685 117515 Phytoene dehydrogenase (Phytoene desaturase) 142 AAZ73149 72536084 phytoene desaturase [Enterobacteriaceae bacterium DC413] 143 ZP_016 . . . 119476622 phytoene dehydrogenase [marine gamma proteobacterium HTCC2143] 144 ABL97778 119713729 crtI phytoene dehydrogenase [uncultured marine bacterium HF10_29C11] 145 ABL97829 119713781 crtI/crtB [uncultured marine bacterium HF10_49E08] 146 EDB02259 142846369 hypothetical protein GOS_1894993 [marine metagenome] 147 ECQ63324 141245988 hypothetical protein GOS_4882732 [marine metagenome] 148 EAS48195 90333025 phytoene dehydrogenase [marine gamma proteobacterium HTCC2207] 149 EAU13549 113732481 Amine oxidase: FAD dependent oxidoreductase [Caulobacter sp. K31] 150 ECV72481 142118377 hypothetical protein GOS_2846587 [marine metagenome] 151 ECZ41342 142625145 hypothetical protein GOS_2188322 [marine metagenome] 152 EDF61582 143556399 hypothetical protein GOS_924048 [marine metagenome] 153 EBN79105 136382241 hypothetical protein GOS_8187951 [marine metagenome] 154 YP_001 . . . 156974447 hypothetical protein VIBHAR_02162 [Vibrio harveyi ATCC BAA-1116] 155 BAA20276 2130971 phytoene desaturase [Erythrobacter longus] 156 EBC81369 134685202 hypothetical protein GOS_10270 [marine metagenome] 157 ECS97023 141726243 hypothetical protein GOS_8923124 [marine metagenome] 158 ECY38926 142480300 hypothetical protein GOS_2368541 [marine metagenome] 159 YP_632230 108758926 putative phytoene dehydrogenase [Myxococcus xanthus DK 1622] 160 EDB72930 142948834 hypothetical protein GOS_1603748 [marine metagenome] 161 EDJ54243 144192357 hypothetical protein GOS_1677113 [marine metagenome] 162 EAR55130 89049570 phytoene dehydrogenase [Photobacterium sp. SKA34] 163 EDF01226 143447606 hypothetical protein GOS_1030671 [marine metagenome] 164 EDD12651 143142675 hypothetical protein GOS_1354831 [marine metagenome] 165 EDI08258 143990952 hypothetical protein GOS_491661 [marine metagenome] 166 EDI22441 144010457 hypothetical protein GOS_467843 [marine metagenome] 167 ABF54837 98978686 amine oxidase [Sphingopyxis alaskensis RB2256] 168 EBL74107 136074962 hypothetical protein GOS_8521797 [marine metagenome] 169 EBX98247 138142760 hypothetical protein GOS_6427327 [marine metagenome] 170 ECV76618 142124100 hypothetical protein GOS_2839194 [marine metagenome] 171 EBP18308 136596794 hypothetical protein GOS_7953009 [marine metagenome] 172 YP_265547 71082828 phytoene dehydrogenase [Candidatus Pelagibacter ubique HTCC1062] 173 EAS85201 91718551 phytoene dehydrogenase [Candidatus Pelagibacter ubique HTCC1002] 174 EDG91241 143827295 hypothetical protein GOS_699208 [marine metagenome] 175 ECV24805 142059343 hypothetical protein GOS_2935833 [marine metagenome] 176 AAY87313 68164584 predicted phytoene desaturase [uncultured bacterium BAC17H8] 177 EBD43768 134788236 hypothetical protein GOS_9929916 [marine metagenome] 178 EDG35515 143701587 hypothetical protein GOS_796054 [marine metagenome] 179 ZP_018 . . . 149185744 Phytoene dehydrogenase [Erythrobacter sp. SD-21] 180 ABL60986 119094156 phytoene dehydrogenase CrtI [uncultured marine bacterium HF10_19P19] 181 ECZ18273 142592252 hypothetical protein GOS_2227647 [marine metagenome] 182 ABD26256 87135514 amine oxidase [Novosphingobium aromaticivorans DSM 12444] 183 EDG45712 143723898 hypothetical protein GOS_778645 [marine metagenome] 184 EBE76519 134988930 hypothetical protein GOS_9708797 [marine metagenome] 185 EDC75856 143092398 hypothetical protein GOS_1419598 [marine metagenome] 186 EDF35075 143504771 hypothetical protein GOS_970643 [marine metagenome] 187 ZP_014 . . . 114771897 phytoene dehydrogenase [alpha proteobacterium HTCC2255] 188 EDG12368 143659594 hypothetical protein GOS_836451 [marine metagenome] 189 AAT90320 50952787 putative phytoene dehydrogenase [uncultured proteobacterium eBACred25D05] 190 EBV02567 137608266 hypothetical protein GOS_6968229 [marine metagenome] 191 ECB29435 138684670 hypothetical protein GOS_4941392 [marine metagenome] 192 EDI67354 144073017 hypothetical protein GOS_393156 [marine metagenome] 193 AAY78594 67633343 predicted phytoene dehydrogenase [uncultured bacterium MedeBAC82F10] 194 ECY07797 142438219 hypothetical protein GOS_2421570 [marine metagenome] 195 EDI16519 144002234 hypothetical protein GOS_477455 [marine metagenome] 196 EDH58504 143920724 hypothetical protein GOS_579102 [marine metagenome] 197 EAU52387 114549505 phytoene dehydrogenase CrtI [alpha proteobacterium HTCC2255] 198 ECX42733 142346738 hypothetical protein GOS_2540243 [marine metagenome] 199 EBE06505 134884139 hypothetical protein GOS_9826927 [marine metagenome] 200 EDB92338 142976670 hypothetical protein GOS_1568269 [marine metagenome] 201 EBN30363 136310418 hypothetical protein GOS_8268666 [marine metagenome]

TABLE 18 Examples of phytoene synthase and lycopene cyclase polypeptides. Accession GI Number Number Description 1613414C 227040 crtB gene A49558 1076590 phytoene synthase 2 precursor - tomato (fragment) AAA19428 506623 phytoene synthase [Neurospora crassa] AAA32836 413732 phytoene synthase AAA64982 148413 phytoene synthase [Pantoea agglomerans] AAB87738 29893495 phytoene synthase [Rubrivivax gelatinosus] AAC44849 1842243 phytoene synthase AAD38051 13542332 phytoene synthase [Citrus × paradisi] AAF78202 8650418 phytoene synthase [Bradyrhizobium sp. ORS278] AAF82616 9081847 phytoene synthase [Tagetes erecta] AAG10427 9971814 phytoene synthase [Tagetes erecta] AAG28701 11066678 CrtB [Streptomyces griseus] AAK07734 18476085 phytoene synthase [Oryza sativa (japonica cultivar-group)] AAK07735 18476089 phytoene synthase [Oryza sativa (japonica cultivar-group)] AAK15621 13195243 phytoene synthase [Haematococcus pluvialis] AAL02001 15553721 phytoene synthase [Xanthobacter sp. Py2] AAL76346 18645045 phytoene synthase [uncultured proteobacterium] AAL82578 21326700 phytoene synthase radicle isoform [Oryza sativa (indica cultivar- group)] AAM45379 21360353 phytoene synthase [Tagetes erecta] AAM48647 21328641 phytoene synthase [uncultured proteobacterium] AAM62787 21553694 phytoene synthase [Arabidopsis thaliana] AAM94363 22296799 phytoene synthase [Agromyces mediolanus] AAN85600 27228294 Phytoene Synthase [Pantoea stewartii] AAO24767 27903500 phytoene synthase [Citrus maxima] AAO39835 28403302 phytoene synthase [Citrus sinensis] AAO46895 37729028 lycopene cyclase/phytoene synthase [Blakeslea trispora] AAO47570 33465823 phytoene-beta carotene synthase [Xanthophyllomyces dendrorhous] AAO73816 33465821 crtYB [Xanthophyllomyces dendrorhous] AAP22038 30349414 phytoene synthase 2 [Zea mays] AAP55451 32350232 phytoene synthase 2 [Zea mays] AAP55453 32350236 phytoene synthase 2 [Zea mays] AAP55461 32350252 phytoene synthase 2 [Zea mays] AAP55471 32350272 phytoene synthase 2 [Zea mays] AAP55484 32350298 phytoene synthase 2 [Zea mays] AAP55486 32350302 phytoene synthase 2 [Zea mays] AAP56083 32349564 phytoene synthase [Zea mays] AAP56124 32349646 phytoene synthase [Zea mays] AAP56127 32349652 phytoene synthase [Zea mays] AAP56136 32349670 phytoene synthase [Zea mays] AAP56148 32349694 phytoene synthase [Zea mays] AAP56155 32349708 phytoene synthase [Zea mays] AAP56156 32349710 phytoene synthase [Zea mays] AAP56157 32349712 phytoene synthase [Zea mays] AAP56158 32349714 phytoene synthase [Zea mays] AAP79176 32307542 phytoene synthase [Bigelowiella natans] AAQ91837 37499616 phytoene synthase 2 [Zea mays] AAR08445 38037628 chloroplast phytoene synthase 1 [Zea mays] AAR31885 39842609 chloroplast phytoene synthase [Zea mays] AAR37803 40062932 phytoene synthase [uncultured bacterium 442] AAR37856 40062986 phytoene synthase [uncultured bacterium 443] AAR86104 40456029 phytoene synthase [Momordica charantia var. abbreviata] AAR87868 40557193 phytoene synthase [Oncidium cv. ‘Gower Ramsey’] AAR98492 41018901 phytoene synthase [Bradyrhizobium sp. ORS278] AAS02284 41394357 phytoene synthase 2 [Zea mays] AAS17009 42491736 phytoene synthase 2 [Oryza sativa (japonica cultivar-group)] AAS18307 42521626 phytoene synthase 1; PSY1 [Oryza sativa (indica cultivar-group)] AAT28184 47498515 phytoene synthase [Dunaliella salina] AAT35222 47531118 fusion of carotene synthesis proteins [synthetic construct] AAT38473 47779181 chloroplast phytoene synthase precursor [Chlamydomonas reinhardtii] AAT46069 48686711 phytoene synthase [Dunaliella salina] AAT74581 50313418 PSY [Citrus sinensis] AAT90319 50952786 putative phytoene synthase [uncultured proteobacterium eBACred25D05] AAV74394 56122551 phytoene synthase [Adonis palaestina] AAW23162 56698928 phytoene synthase [Rhodococcus erythropolis] AC2035 25366683 phytoene synthase [imported] - Nostoc sp. (strain PCC 7120) AC2035 25366683 phytoene synthase [imported] - Nostoc sp. (strain PCC 7120) BAB18514 11344507 phytoene synthase [Citrus unshiu] BAB79604 18143449 crtB [Pantoea agglomerans pv. milletiae] BAD07278 40809739 phytoene synthase [Citrus sinensis] BAD07286 40809755 phytoene synthase [Citrus limon] BAD62106 54291340 phytoene synthase [Oryza sativa (japonica cultivar-group)] BAD62107 54291341 phytoene synthase-like [Oryza sativa (japonica cultivar-group)] C90061 25506636 squalene desaturase [imported] - Staphylococcus aureus (strain N315) CAA47625 19347 mutant phytoene synthase [Lycopersicon esculentum] CAA68575 19341 unnamed protein product [Lycopersicon esculentum] CAB07958 1934837 unknown [Bacillus subtilis] CAB38740 4490590 phytoene synthase [Rhodobacter sphaeroides] CAB51949 5690074 phytoene-beta carotene synthase [Xanthophyllomyces dendrorhous] CAB56063 5912296 phytoene synthase [Paracoccus marcusii] CAB86388 7453011 phytoene synthase/lycopene cyclase [Phycomyces blakesleeanus] CAB93661 8250520 lycopene cyclase/phytoene synthase [Phycomyces blakesleeanus] CAB94795 8574392 phytoene synthase [Mycobacterium aurum] CAC19567 11990226 phytoene synthase [Helianthus annuus] CAC27383 12584564 phytoene synthase [Helianthus annuus] CAD19988 18307500 carotene cyclase [Gibberella fujikuroi] CAD29284 57282088 putative phytoene synthase [Oryza sativa] CAE76609 38567321 geranylgeranyl-diphosphate geranylgeranyltransferase (AL-2) [Neurospora crassa] E37802 95606 crtB protein - Erwinia uredovora E84320 25410251 phytoene synthase [imported] - Halobacterium sp. NRC-1 EAA98758 42863045 unknown [environmental sequence] EAB01965 42869439 unknown [environmental sequence] EAB04170 42873822 unknown [environmental sequence] EAB07138 42879858 unknown [environmental sequence] EAB09791 42885235 unknown [environmental sequence] EAB19826 42905452 unknown [environmental sequence] EAB35029 42936011 unknown [environmental sequence] EAB41375 42948740 unknown [environmental sequence] EAB78706 43024004 unknown [environmental sequence] EAB92586 43052355 unknown [environmental sequence] EAC06949 43081493 unknown [environmental sequence] EAC18360 43104624 unknown [environmental sequence] EAC25793 43119723 unknown [environmental sequence] EAC29883 43128092 unknown [environmental sequence] EAC32813 43133973 unknown [environmental sequence] EAC33105 43134560 unknown [environmental sequence] EAC38486 43145552 unknown [environmental sequence] EAC52233 43173313 unknown [environmental sequence] EAC60029 43189028 unknown [environmental sequence] EAC68026 43204953 unknown [environmental sequence] EAC96197 43261031 unknown [environmental sequence] EAD08701 43285745 unknown [environmental sequence] EAD20866 43310220 unknown [environmental sequence] EAD32755 43334458 unknown [environmental sequence] EAD38008 43345761 unknown [environmental sequence] EAD50152 43370658 unknown [environmental sequence] EAD50402 43371147 unknown [environmental sequence] EAD81123 43452903 unknown [environmental sequence] EAD93882 43478303 unknown [environmental sequence] EAE12860 43516265 unknown [environmental sequence] EAE16121 43522884 unknown [environmental sequence] EAE31084 43552634 unknown [environmental sequence] EAE35665 43561764 unknown [environmental sequence] EAE44717 43579862 unknown [environmental sequence] EAE46627 43583580 unknown [environmental sequence] EAE47846 43586023 unknown [environmental sequence] EAE72264 43635190 unknown [environmental sequence] EAE76009 43642573 unknown [environmental sequence] EAE86335 43662748 unknown [environmental sequence] EAE89581 43669163 unknown [environmental sequence] EAF18881 43728007 unknown [environmental sequence] EAF64277 43819669 unknown [environmental sequence] EAF67931 43827263 unknown [environmental sequence] EAF84745 43861327 unknown [environmental sequence] EAF94004 43880040 unknown [environmental sequence] EAG06083 43903395 unknown [environmental sequence] EAG21950 43933540 unknown [environmental sequence] EAG43625 43973477 unknown [environmental sequence] EAG50171 43985555 unknown [environmental sequence] EAG57517 43999205 unknown [environmental sequence] EAG62787 44009110 unknown [environmental sequence] EAG65580 44014171 unknown [environmental sequence] EAG68110 44018715 unknown [environmental sequence] EAG72283 44026322 unknown [environmental sequence] EAG78750 44037938 unknown [environmental sequence] EAG80445 44041116 unknown [environmental sequence] EAG93220 44064453 unknown [environmental sequence] EAH04927 44085694 unknown [environmental sequence] EAH08972 44093217 unknown [environmental sequence] EAH10377 44095788 unknown [environmental sequence] EAH22151 44117864 unknown [environmental sequence] EAH31461 44134654 unknown [environmental sequence] EAH50033 44169323 unknown [environmental sequence] EAH64480 44196848 unknown [environmental sequence] EAH79040 44223009 unknown [environmental sequence] EAH99976 44255671 unknown [environmental sequence] EAI02786 44259828 unknown [environmental sequence] EAI02787 44259829 unknown [environmental sequence] EAI03575 44260943 unknown [environmental sequence] EAI05900 44264266 unknown [environmental sequence] EAI61004 44344824 unknown [environmental sequence] EAI70669 44358327 unknown [environmental sequence] EAI83938 44377067 unknown [environmental sequence] EAJ05110 44406802 unknown [environmental sequence] EAJ05569 44407471 unknown [environmental sequence] EAJ08876 44412338 unknown [environmental sequence] EAJ35156 44449986 unknown [environmental sequence] EAJ38900 44455130 unknown [environmental sequence] EAJ49645 44470504 unknown [environmental sequence] EAJ54357 44477026 unknown [environmental sequence] EAJ60475 44485647 unknown [environmental sequence] EAJ64125 44492007 unknown [environmental sequence] EAJ67499 44497025 unknown [environmental sequence] EAJ76471 44510405 unknown [environmental sequence] EAJ76950 44511114 unknown [environmental sequence] EAJ78637 44513596 unknown [environmental sequence] EAJ78787 44513824 unknown [environmental sequence] EAJ79616 44515082 unknown [environmental sequence] EAJ80356 44516200 unknown [environmental sequence] EAJ81914 44518489 unknown [environmental sequence] EAJ87417 44526623 unknown [environmental sequence] EAK08514 44557109 unknown [environmental sequence] EAK08523 44557119 unknown [environmental sequence] EAK12901 44563097 unknown [environmental sequence] EAK22180 44576315 unknown [environmental sequence] EAK24859 44580262 unknown [environmental sequence] EAK28345 44585276 unknown [environmental sequence] EAK34732 44594324 unknown [environmental sequence] EAK34736 44594329 unknown [environmental sequence] EAK37296 44597942 unknown [environmental sequence] EAK37521 44598256 unknown [environmental sequence] EAK56335 44624430 unknown [environmental sequence] G84363 25410528 hypothetical protein ggt [imported] - Halobacterium sp. NRC-1 NP_274195 15677043 phytoene synthase, putative [Neisseria meningitidis MC58] NP_284085 15794263 poly-isoprenyl transferase [Neisseria meningitidis Z2491] NP_294586 15805888 phytoene synthase [Deinococcus radiodurans R1] NP_388961 16078144 hypothetical protein BSU10810 [Bacillus subtilis subsp. subtilis str. 168] NP_441168 16330440 phytoene synthase [Synechocystis sp. PCC 6803] NP_443763 16519643 Y4aC [Rhizobium sp. NGR234] NP_624523 21218744 putative phytoene synthase [Streptomyces coelicolor A3(2)] NP_630832 21225053 putative phytoene synthase [Streptomyces coelicolor A3(2)] NP_662273 21674208 phytoene desaturase [Chlorobium tepidum TLS] NP_682350 22299103 phytoene synthase [Thermosynechococcus elongatus BP-1] NP_693381 23099915 phytoene synthase [Oceanobacillus iheyensis HTE831] NP_786525 28379633 phytoene synthase [Lactobacillus plantarum WCFS1] NP_822199 29827565 phytoene synthase [Streptomyces avermitilis MA-4680] NP_822829 29828195 squalene/phytoene synthase [Streptomyces avermitilis MA-4680] NP_851527 30795077 putative phytoene synthase [Streptomyces rochei] NP_868799 32475805 probable phytoene synthase [Rhodopirellula baltica SH 1] NP_874560 33239618 Phytoene synthase, CrtB [Prochlorococcus marinus subsp. marinus str. CCMP1375] NP_879992 33592348 putative phytoene synthase [Bordetella pertussis Tohama I] NP_884101 33596458 putative phytoene synthase [Bordetella parapertussis 12822] NP_889809 33602249 putative phytoene synthase [Bordetella bronchiseptica RB50] NP_892264 33860703 Squalene and phytoene synthases [Prochlorococcus marinus subsp. pastoris str. CCMP1986] NP_895828 33864268 Squalene and phytoene synthases [Prochlorococcus marinus str. MIT 9313] NP_898345 33866786 phytoene synthases [Synechococcus sp. WH 8102] NP_902648 34498433 probable geranylgeranyl-diphosphate geranylgeranyltransferase [Chromobacterium violaceum ATCC 12472 NP_902649 34498434 probable phytoene synthase-related protein [Chromobacterium violaceum ATCC 12472] NP_924690 37521313 phytoene synthase [Gloeobacter violaceus PCC 7421] NP_931516 37528171 phytoene synthase [Photorhabdus luminescens subsp. laumondii TTO1] NP_946861 39934585 CrtB phytoene synthase [Rhodopseudomonas palustris CGA009] NP_949079 39936803 putative poly-isoprenyl transferase [Rhodopseudomonas palustris CGA009] NP_962005 41409169 hypothetical protein MAP3071 [Mycobacterium avium subsp. paratuberculosis str. k10] NP_968601 42523221 phytoene synthase [Bdellovibrio bacteriovorus HD100] O07333 3913360 Phytoene synthase P08196 585746 Phytoene synthase 1, chloroplast precursor (Fruit ripening specific protein pTOM5) P21683 30923192 Phytoene synthase P37269 585009 Phytoene synthase P37271 27735222 Phytoene synthase, chloroplast precursor P37272 585749 Phytoene synthase, chloroplast precursor P53797 1709885 Phytoene synthase, chloroplast precursor P54975 1706137 Phytoene synthase P54977 1706139 Phytoene synthase P65860 54041032 Probable phytoene synthase Q9SSU8 8928282 Phytoene synthase, chloroplast precursor Q9UUQ6 34922667 CarRP protein [Includes: Lycopene cyclase; Phytoene synthase] S22474 7489041 phytoene synthase (EC 2.5.1.—) - tomato S32170 321671 phytoene synthetase - Myxococcus xanthus S52587 1073300 prephytoene pyrophosphate synthase - Erwinia herbicola S56668 2129505 geranylgeranyl-diphosphate geranylgeranyltransferase (EC 2.5.1.32) precursor - muskmelon S68307 2130144 phytoene synthase - maize T10702 7484346 phytoene synthase (EC 2.5.1.—) - green alga (Dunaliella bardawil) T46594 11291807 phytoene synthase (EC 2.5.1.—) [validated] - Mycobacterium marinum T50746 11356347 phytoene synthase (EC 2.5.1.—) [imported] - Rhodobacter sphaeroides T50895 11291816 prephytoene pyrophosphate synthase [imported] - Rubrivivax gelatinosus XP_324765 32408567 PHYTOENE SYNTHASE [Neurospora crassa] XP_383242 46114448 hypothetical protein FG03066.1 [Gibberella zeae PH-1] XP_403902 49080862 hypothetical protein UM06287.1 [Ustilago maydis 521] YP_006040 46255128 phytoene synthase [Thermus thermophilus HB27] YP_103126 53723680 phytoene synthase, putative [Burkholderia mallei ATCC 23344] YP_112342 53723357 squalene/phytoene synthase [Burkholderia pseudomallei K96243] YP_117945 54023703 putative phytoene synthase [Nocardia farcinica IFM 10152] YP_120611 54026369 putative phytoene synthase [Nocardia farcinica IFM 10152] YP_136628 55378778 lycopene cyclase [Haloarcula marismortui ATCC 43049] YP_136629 55378779 phytoene synthase [Haloarcula marismortui ATCC 43049] YP_145340 55978284 phytoene synthase [Thermus thermophilus HB8] YP_145343 55978287 phytoene synthase-related protein [Thermus thermophilus HB8] YP_160917 56479328 probable terpenoid synthase-related protein [Azoarcus sp. EbN1] YP_160918 56479329 putative terpenoid synthase [Azoarcus sp. EbN1] YP_162605 56551766 phytoene/squalene synthetase [Zymomonas mobilis subsp. mobilis ZM4] YP_172822 56752121 phytoene synthase [Synechococcus elongatus PCC 6301] YP_187369 57652299 dehydrosqualene synthase [Staphylococcus aureus subsp. aureus COL] YP_192648 58040684 Putative phytoene synthase [Gluconobacter oxydans 621H] ZP_000044 22956752 COG1562: Phytoene/squalene synthetase [Rhodobacter sphaeroides 2.4.1] ZP_001091 53688068 COG1562: Phytoene/squalene synthetase [Nostoc punctiforme PCC 73102] ZP_001591 53763709 COG1562: Phytoene/squalene synthetase [Anabaena variabilis ATCC 29413] ZP_001657 45514234 COG1562: Phytoene/squalene synthetase [Ralstonia eutropha JMP134] ZP_001690 46132223 COG1562: Phytoene/squalene synthetase [Ralstonia eutropha JMP134] ZP_001746 45523280 COG1562: Phytoene/squalene synthetase [Crocosphaera watsonii WH 8501] ZP_001837 53771530 COG1562: Phytoene/squalene synthetase [Exiguobacterium sp. 255- 15] ZP_001867 45546711 COG1562: Phytoene/squalene synthetase [Rubrobacter xylanophilus DSM 9941] ZP_002096 46204978 COG1562: Phytoene/squalene synthetase [Magnetospirillum magnetotacticum MS-1] ZP_002248 46324513 COG1562: Phytoene/squalene synthetase [Burkholderia cepacia R1808] ZP_002450 47575031 COG1562: Phytoene/squalene synthetase [Rubrivivax gelatinosus PM1] ZP_002680 48763469 COG1562: Phytoene/squalene synthetase [Rhodospirillum rubrum] ZP_002710 48766450 COG1562: Phytoene/squalene synthetase [Rhodospirillum rubrum] ZP_002791 48782680 COG1562: Phytoene/squalene synthetase [Burkholderia fungorum LB400] ZP_002892 48832182 COG1562: Phytoene/squalene synthetase [Magnetococcus sp. MC-1] ZP_002916 48834623 COG1562: Phytoene/squalene synthetase [Thermobifida fusca] ZP_003036 48849426 COG1562: Phytoene/squalene synthetase [Novosphingobium aromaticivorans DSM 12444] ZP_003269 48893702 COG1562: Phytoene/squalene synthetase [Trichodesmium erythraeum IMS101] ZP_003351 52007802 COG1562: Phytoene/squalene synthetase [Thiobacillus denitrificans ATCC 25259] ZP_003487 53730362 COG1562: Phytoene/squalene synthetase [Dechloromonas aromatica RCB] ZP_003501 53759405 COG1562: Phytoene/squalene synthetase [Methylobacillus flagellatus KT] ZP_003591 53798896 COG1562: Phytoene/squalene synthetase [Chloroflexus aurantiacus] ZP_003628 54030691 COG1562: Phytoene/squalene synthetase [Polaromonas sp. JS666]

TABLE 19 Examples of carotenoid ketolase polypeptides. Accession GI Number Number Description AAA99932 609575 fatty acid desaturase AAB48668 1870215 ORF [Emericella nidulans] AAC25611 2541936 unknown [Pseudomonas syringae] AAF78203 8650419 beta-carotene ketolase [Bradyrhizobium sp. ORS278] AAH16427 16741158 2210008A03Rik protein [Mus musculus] AAN03484 22597194 BKT [Haematococcus pluvialis] AAN85497 26541510 putative fatty acid desaturase [Streptomyces atroolivaceus] AAN86030 33439708 beta-carotene C4 oxygenase [Brevundimonas aurantiaca] AAO64399 28976134 putative beta-carotene ketolase [Nodularia spumigena] AAQ23139 33621091 CrtW/CrtY fusion protein [synthetic construct] AAT35222 47531118 fusion of carotene synthesis proteins [synthetic construct] AAT35555 47558911 beta-carotene ketolase [Haematococcus pluvialis] AAV41372 55139370 beta-carotene ketolase/oxygenase [‘Chlorella’ zofingiensis] AB2307 25530134 hypothetical protein alr4009 [imported] - Nostoc sp. (strain PCC 7120) AF2204 25533132 beta-carotene ketolase [imported] - Nostoc sp. (strain PCC 7120) BAB54999 14028447 mlr9395 [Mesorhizobium loti MAFF303099] BAB58879 14270087 membrane fatty acid desaturase [Toxoplasma gondii] BAC98366 37360914 alkane hydroxylase [Alcanivorax borkumensis] CAA60478 2654318 beta-carotene C-4 oxygenase (ketolase) [Haematococcus pluvialis] CAB56059 5912292 beta-carotene C-4-oxygenase (ketolase) [Paracoccus marcusii] D87673 25398945 conserved hypothetical protein CC3422 [imported] - Caulobacter crescentus EAA79304 42823978 unknown [environmental sequence] EAA80363 42826055 unknown [environmental sequence] EAA81403 42828115 unknown [environmental sequence] EAA84711 42834481 unknown [environmental sequence] EAB82380 43031476 unknown [environmental sequence] EAB86624 43040184 unknown [environmental sequence] EAC05755 43079085 unknown [environmental sequence] EAD12219 43292778 unknown [environmental sequence] EAD71182 43427899 unknown [environmental sequence] EAD94927 43480380 unknown [environmental sequence] EAF11582 43712986 unknown [environmental sequence] EAF98072 43888329 unknown [environmental sequence] EAG19345 43928738 unknown [environmental sequence] EAG38273 43963688 unknown [environmental sequence] EAG79800 44039853 unknown [environmental sequence] EAG96474 44070318 unknown [environmental sequence] EAH00349 44077315 unknown [environmental sequence] EAH36448 44143633 unknown [environmental sequence] EAH40683 44151265 unknown [environmental sequence] EAH53180 44175316 unknown [environmental sequence] EAH96648 44250729 unknown [environmental sequence] EAI05260 44263397 unknown [environmental sequence] EAI17468 44281329 unknown [environmental sequence] EAI53009 44333409 unknown [environmental sequence] EAI54054 44334878 unknown [environmental sequence] EAI67818 44354362 unknown [environmental sequence] EAI68153 44354875 unknown [environmental sequence] EAI89684 44384943 unknown [environmental sequence] EAJ27674 44439188 unknown [environmental sequence] EAJ45589 44464684 unknown [environmental sequence] EAJ45589 44464684 unknown [environmental sequence] EAJ67118 44496466 unknown [environmental sequence] EAJ74221 44507022 unknown [environmental sequence] EAJ74653 44507662 unknown [environmental sequence] EAJ88396 44528064 unknown [environmental sequence] EAJ88887 44528792 unknown [environmental sequence] EAK06069 44553531 unknown [environmental sequence] EAK11467 44561166 unknown [environmental sequence] EAK16824 44568733 unknown [environmental sequence] EAK28828 44585942 unknown [environmental sequence] EAK28828 44585942 unknown [environmental sequence] EAK31112 44589271 unknown [environmental sequence] EAK42591 44605441 unknown [environmental sequence] NP_045063 11465545 fatty-acid desaturase [Cyanidium caldarium] NP_081575 27754029 sphingolipid delta 4 desaturase/C-4 hydroxylase [Mus musculus] NP_338204 15843167 fatty acid desaturase, putative/ferredoxin reductase, electron transfer component [Mycobacterium tuberculosis CDC1551] NP_440788 16330060 b-carotene hydroxylase [Synechocystis sp. PCC 6803] NP_441220 16330492 hypothetical protein sl11611 [Synechocystis sp. PCC 6803] NP_682690 22299443 beta-carotene hydroxylase [Thermosynechococcus elongatus BP-1] NP_770721 27379192 hypothetical protein blr4081 [Bradyrhizobium japonicum USDA 110] NP_848964 30468077 beta-carotene hydroxylase [Cyanidioschyzon merolae strain 10D] NP_857223 31794730 POSSIBLE ELECTRON TRANSFER PROTEIN FDXB [Mycobacterium bovis AF2122/97] NP_881760 33594116 putative fatty acid desaturase [Bordetella pertussis Tohama I] NP_882469 33594826 putative fatty acid desaturase [Bordetella parapertussis 12822] NP_886657 33599097 putative fatty acid desaturase [Bordetella bronchiseptica RB50] NP_895643 33864083 beta carotene hydroxylase [Prochlorococcus marinus str. MIT 9313] NP_896386 33864827 beta-carotene hydroxylase [Synechococcus sp. WH 8102] NP_897461 33865902 possible beta-carotene ketolase [Synechococcus sp. WH 8102] NP_924674 37521297 beta-carotene ketolase [Gloeobacter violaceus PCC 7421] NP_927525 37524181 hypothetical protein plu0159 [Photorhabdus luminescens subsp. laumondii TTO1] NP_947075 39934799 Fatty acid desaturase family [Rhodopseudomonas palustris CGA009] P54972 1706150 Beta-carotene ketolase (Beta-carotene oxygenase) Q39982 2498257 Beta-carotene ketolase (Beta-carotene oxygenase) Q44261 2498256 Beta-carotene ketolase (Beta-carotene oxygenase) T31123 11361063 hypothetical protein 15 - Sphingomonas aromaticivorans plasmid pNL1 XP_330780 32420673 hypothetical protein [Neurospora crassa] XP_368852 39974923 hypothetical protein MG00392.4 [Magnaporthe grisea 70-15] XP_380194 46102628 hypothetical protein FG00018.1 [Gibberella zeae PH-1] XP_383758 46115480 hypothetical protein FG03582.1 [Gibberella zeae PH-1] XP_405100 49086048 hypothetical protein AN0963.2 [Aspergillus nidulans FGSC A4] XP_409222 49095522 hypothetical protein AN5085.2 [Aspergillus nidulans FGSC A4] YP_102417 53725671 alkane-1 monooxygenase [Burkholderia mallei ATCC 23344] YP_108945 53719959 putative alkane monooxygenase [Burkholderia pseudomallei K96243] YP_132414 54302421 hypothetical putative delta-9 fatty acid desaturase [Photobacterium profundum SS9] YP_154670 56459389 Fatty-acid desaturase [Idiomarina loihiensis L2TR] YP_166682 56696325 fatty acid desaturase family protein [Silicibacter pomeroyi DSS-3] YP_168846 56698471 fatty acid desaturase family protein [Silicibacter pomeroyi DSS-3] YP_172377 56751676 beta-carotene hydroxylase [Synechococcus elongatus PCC 6301] ZP_001068 23124870 COG3239: Fatty acid desaturase [Nostoc punctiforme PCC 73102] ZP_001112 53688676 COG3239: Fatty acid desaturase [Nostoc punctiforme PCC 73102] ZP_001607 53764743 COG3239: Fatty acid desaturase [Anabaena variabilis ATCC 29413] ZP_001757 46118877 COG3239: Fatty acid desaturase [Crocosphaera watsonii WH 8501] ZP_001787 53736018 COG3239: Fatty acid desaturase [Crocosphaera watsonii WH 8501] ZP_002218 46321435 COG3239: Fatty acid desaturase [Burkholderia cepacia R1808] ZP_002456 47575608 COG3239: Fatty acid desaturase [Rubrivivax gelatinosus PM1] ZP_003028 48848557 COG3239: Fatty acid desaturase [Novosphingobium aromaticivorans DSM 12444] ZP_003107 48856640 COG3239: Fatty acid desaturase [Cytophaga hutchinsonii] ZP_003264 48893204 COG3239: Fatty acid desaturase [Trichodesmium erythraeum IMS101] ZP_003458 53688805 hypothetical protein Npun02000865 [Nostoc punctiforme PCC 73102] ZP_003513 53763576 COG3239: Fatty acid desaturase [Anabaena variabilis ATCC 29413]

TABLE 20 Examples of carotenoid hydroxylase polypeptides. Accession GI Number Number Description AAC44852 1842246 b-carotene hydroxylase AAC49443 1575296 beta-carotene hydroxylase [Arabidopsis thaliana] AAD54243 5852870 carotenoid hydroxylase [Haematococcus pluvialis] AAG10430 9971820 beta hydroxylase [Tagetes erecta] AAG10793 9988836 beta-carotene hydroxylase [Citrus unshiu] AAG33636 11245486 beta-carotene hydroxylase [Citrus unshiu] AAL80006 19071768 beta carotene hydroxylase [Sandersonia aurantiaca] AAM44971 21280903 putative beta-carotene hydroxylase [Arabidopsis thaliana] AAM51300 21436107 putative beta-carotene hydroxylase [Arabidopsis thaliana] AAM77007 21734857 beta-carotene hydroxylase [Vitis vinifera] AAN85601 27228295 Beta-Carotene Hydroxylase [Pantoea stewartii] AAO53295 28911949 carotenoid hydroxylase [Haematococcus pluvialis] AAS48097 44887642 beta-carotene hydroxylase [Citrus sinensis] AAS55552 45184599 putative beta-carotene hydroxylase [Brassica rapa] AAS88426 46326968 beta-carotene hydroxylase [Glycine max] AAT48741 49036137 beta-carotene hydroxylase [Citrus sinensis] AAT84408 50844570 beta carotene hydroxylase [Crocus sativus] AAV85452 56267980 beta-ring oxygenase AdKeto1 [Adonis palaestina] AAV85453 56267982 beta-ring oxygenase AdKeto2 [Adonis palaestina] BAA14129 216687 crtZ [Pantoea ananatis] BAB79605 18143450 crtZ [Pantoea agglomerans pv. milletiae] BAC77670 31790567 gamma-carotene hydroxylase [marine bacterium P99-3] BAD07283 40809749 beta-ring hydroxylase [Citrus sinensis] BAD07291 40809765 beta-ring hydroxylase [Citrus limon] CAA70427 2956671 beta-carotene hydrolase [Capsicum annuum] CAA70888 2956717 beta-carotene hydroxylase 2 [Capsicum annuum] CAB55625 5870598 beta-carotene hydroxylase [Lycopersicon esculentum] CAB55626 5870600 beta-carotene hydroxylase [Lycopersicon esculentum] CAB56060 5912293 carotene hydroxylase [Paracoccus marcusii] CAC06712 9968545 beta-carotene hydroxylase [Narcissus pseudonarcissus] CAC95130 33145986 beta-carotene hydroxylase [Crocus sativus] EAB30128 42926157 unknown [environmental sequence] EAC49462 43167766 unknown [environmental sequence] EAC86129 43241003 unknown [environmental sequence] EAD61089 43395962 unknown [environmental sequence] EAD76156 43443111 unknown [environmental sequence] EAD88640 43467793 unknown [environmental sequence] EAE27903 43546376 unknown [environmental sequence] EAE28203 43546980 unknown [environmental sequence] EAE78743 43647896 unknown [environmental sequence] EAF12173 43714211 unknown [environmental sequence] EAH29370 44130906 unknown [environmental sequence] EAH44202 44158360 unknown [environmental sequence] EAI00766 44256844 unknown [environmental sequence] EAI29017 44298625 unknown [environmental sequence] EAJ30844 44443849 unknown [environmental sequence] EAJ72524 44504516 unknown [environmental sequence] EAK10611 44559981 unknown [environmental sequence] EAK53455 44620561 unknown [environmental sequence] EAK63955 44635271 unknown [environmental sequence] H90469 25394049 beta carotene hydroxylase (crtZ) [imported] - Sulfolobus solfataricus NP_745389 26989964 beta-carotene hydroxylase [Pseudomonas putida KT2440] NP_922503 37536402 putative beta-carotene hydroxylase [Oryza sativa (japonica cultivar-group)] P54973 1706152 Beta-carotene hydroxylase Q44262 2498258 Beta-carotene hydroxylase S52982 1073291 beta-carotene hydroxylase - Erwinia herbicola XP_473611 50928167 OSJNBa0011J08.7 [Oryza sativa (japonica cultivar-group)] YP_024309 48478603 beta carotene hydroxylase [Picrophilus torridus DSM 9790] ZP_003055 48851297 COG3000: Sterol desaturase [Novosphingobium aromaticivorans DSM 12444] ZP_003107 48856620 COG3000: Sterol desaturase [Cytophaga hutchinsonii]

TABLE 21 Examples of astaxanthin synthase polypeptides and putative astaxanthin synthase polypeptides. ACCESSION GI PROTEIN DESCRIPTION AAM56288 21501451 Astaxanthin synthase [Xanthophyllomyces dendrorhous] XP_571276 58268240 Cytochrome P450, putative [Cryptococcus neoformans var. neoformans JEC21] EAL20013 50257304 hypothetical protein CNBF3400 [Cryptococcus neoformans var. neoformans B-3501A] XP_401804 49075484 hypothetical protein UM04189.1 [Ustilago maydis 521] XP_397817 49067054 hypothetical protein UM00202.1 [Ustilago maydis 521] XP_399595 49070612 hypothetical protein UM01980.1 [Ustilago maydis 521] XP_403279 49079218 hypothetical protein UM05664.1 [Ustilago maydis 521] XP_382294 46110473 hypothetical protein FG02118.1 [Gibberella zeae PH-1] XP_406021 49088382 hypothetical protein AN1884.2 [Aspergillus nidulans FGSC A4] XP_381224 46108332 hypothetical protein FG01048.1 [Gibberella zeae PH-1] XP_391479 46139577 hypothetical protein FG11303.1 [Gibberella zeae PH-1] XP_569261 58264210 conserved hypothetical protein [Cryptococcus neoformans var. neoformans JEC21] EAL22841 50260180 hypothetical protein CNBB0620 [Cryptococcus neoformans var. neoformans B-3501A] XP_359866 39940658 hypothetical protein MG04911.4 [Magnaporthe grisea 70-15]

TABLE 22 Examples of carotenoid epsilon hydroxylase polypeptides. ACCESSION GI PROTEIN DESCRIPTION ABB52076 79155148 putative epsilon-ring carotene hydroxylase [Daucus carota subsp. sativus] BAD94136 62319017 Cytochrom P450-like protein [Arabidopsis thaliana] ABD28565 87162770 E-class P450, group I [Medicago truncatula] AAT28222 47498772 putative 97B2-like cytochrome P450 [Ginkgo biloba] ABC68396 85001685 cytochrome P450 monooxygenase CYP97A [Glycine max] ABC59110 84514203 cytochrome P450 monooxygenase CYP97B [Medicago truncatula] NP_190881 42565881 LUT1 (LUTEIN DEFICIENT 1); oxygen binding [Arabidopsis thaliana] ABB47954 78708979 cytochrome P450 monooxygenase, putative [Oryza sativa (japonica cultivar-group)] NP_922604 37536604 putative cytochrome P450 monooxygenase [Oryza sativa (japonica cultivar-group)]

TABLE 23 Examples of lycopene cyclase polypeptides, beta and epsilon subunits. ACCESSION GI PROTEIN DESCRIPTION XP_001422490 145356548 predicted protein [Ostreococcus lucimarinus CCE9901] BAE94036 94481238 lycopene epsilon cyclase (Diospyros kaki ) AAK07431 12746307 lycopene epsilon-cyclase [Adonis palaestina] ABB52073 79154988 putative lycopene epsilon cyclase [Daucus carota subsp. sativus] Q38932 27735211 Lycopene epsilon cyclase, chloroplast precursor AAB53336 1399181 lycopene epsilon cyclase AAG10428 9971816 epsilon cyclase [Tagetes erecta] AAK07434 12746313 lycopene epsilon-cyclase [Lactuca sativa] AAM45382 21360359 epsilon cyclase [Tagetes erecta] O65837 11132841 Lycopene epsilon cyclase, chloroplast precursor AAL69394 18419661 lycopene epsilon-cyclase [Spinacia oleracea] BAE79549 87299433 lycopene epsilon-cyclase [Chrysanthemum × morifolium] XP_463351 50901836 putative lycopene epsilon-cyclase [Oryza sativa (japonica cultivar-group)] AAS48096 44887640 epsilon lycopene cyclase [Citrus sinensis] AAX92679 62638188 lycopene epsilon cyclase [Citrus maxima] AAL92114 19569601 lycopene epsilon-cyclase [Citrus × paradisi] AAK07433 12746311 lycopene epsilon-cyclase [Solanum tuberosum] AAL47019 17864021 lycopene epsilon-cyclase [Citrus sinensis] AAT46065 48686703 chloroplast lycopene epsilon-cyclase precursor [Chlamydomonas reinhardtii] BAD07293 40809769 lycopene epsilon-cyclase [Citrus limon] BAD07285 40809753 lycopene epsilon-cyclase [Citrus sinensis] BAD07277 40809737 lycopene epsilon-cyclase [Citrus unshiu] EAJ62839 44489138 unknown [environmental sequence] BAE43547 73993068 putative lycopene beta cyclase [Taxodium distichum var. distichum] BAE43550 73993074 putative lycopene beta cyclase [Taxodium distichum var. distichum] BAE43557 73993088 putative lycopene beta cyclase [Taxodium distichum var. imbricarium] BAE43558 73993090 putative lycopene beta cyclase [Taxodium distichum var. imbricarium] BAE43553 73993080 putative lycopene beta cyclase [Taxodium distichum var. imbricarium] BAE43545 73993064 putative lycopene beta cyclase [Taxodium distichum var. distichum] BAE43556 73993086 putative lycopene beta cyclase [Taxodium distichum var. imbricarium] BAE43552 73993078 putative lycopene beta cyclase [Taxodium distichum var. distichum] BAE43560 73993094 putative lycopene beta cyclase [Taxodium distichum var. imbricarium] BAE43554 73993082 putative lycopene beta cyclase [Taxodium distichum var. imbricarium] BAE43551 73993076 putative lycopene beta cyclase [Taxodium distichum var. distichum] BAE43519 73993012 putative lycopene beta cyclase [Cryptomeria japonica] BAE43535 73993044 putative lycopene beta cyclase [Cryptomeria japonica] BAE43541 73993056 putative lycopene beta cyclase [Cryptomeria japonica] BAE43542 73993058 putative lycopene beta cyclase [Cryptomeria japonica] BAE43517 73993008 putative lycopene beta cyclase [Cryptomeria japonica] BAE43534 73993042 putative lycopene beta cyclase [Cryptomeria japonica] BAE43537 73993048 putative lycopene beta cyclase [Cryptomeria japonica] BAE43533 73993040 putative lycopene beta cyclase [Cryptomeria japonica] BAD02774 38603277 putative lycopene beta cyclase [Cryptomeria japonica] BAD02766 38603261 putative lycopene beta cyclase [Cryptomeria japonica] BAE43540 73993054 putative lycopene beta cyclase [Cryptomeria japonica] BAE43514 73993002 putative lycopene beta cyclase [Cryptomeria japonica] BAE43544 73993062 putative lycopene beta cyclase [Cryptomeria japonica] BAE43538 73993050 putative lycopene beta cyclase [Cryptomeria japonica] BAE43528 73993030 putative lycopene beta cyclase [Cryptomeria japonica] BAE43546 73993066 putative lycopene beta cyclase [Taxodium distichum var. distichum] BAE43526 73993026 putative lycopene beta cyclase [Cryptomeria japonica] BAE43543 73993060 putative lycopene beta cyclase [Cryptomeria japonica] BAD02742 38603213 putative lycopene beta cyclase [Cryptomeria japonica] BAD02770 38603269 putative lycopene beta cyclase [Cryptomeria japonica] BAE43522 73993018 putative lycopene beta cyclase [Cryptomeria japonica] BAE43559 73993092 putative lycopene beta cyclase [Taxodium distichum var. imbricarium] BAE43527 73993028 putative lycopene beta cyclase [Cryptomeria japonica] BAE43548 73993070 putative lycopene beta cyclase [Taxodium distichum var. distichum] AAF44700 14550425 lycopene beta-cyclase [Citrus sinensis] BAE43555 73993084 putative lycopene beta cyclase [Taxodium distichum var. imbricarium] BAE43549 73993072 putative lycopene beta cyclase [Taxodium distichum var. distichum] AAU14144 51922063 lycopene beta-cyclase [Citrus sinensis] AAN86060 27261727 lycopene cyclase [Citrus unshiu] AAR89632 40756518 lycopene-beta-cyclase [Citrus maxima] AAM21152 20530862 lycopene beta-cyclase [Citrus sinensis] AAD38049 13959731 lycopene cyclase [Citrus × paradisi] AAU05146 51511939 lycopene beta-cyclase [Citrus sinensis] AAU05145 51511937 lycopene beta-cyclase [Citrus sinensis] AAK07430 12746305 lycopene beta-cyclase [Adonis palaestina] ABB72443 82394885 lycopene beta-cyclase [Citrus sinensis] BAE79544 87299423 lycopene beta-cyclase [Chrysanthemum × morifolium] BAE78471 85717882 lycopene beta cyclase [Taraxacum officinale] Q43415 11133019 Lycopene beta cyclase, chloroplast precursor AAF23013 6665782 lycopene epsilon-cyclase [Daucus carota] ABB52071 79154899 putative lycopene beta cyclase [Daucus carota subsp. sativus] AAW88382 59665024 lycopene beta-cyclase [Lycium barbarum] AAG10429 9971818 beta cyclase [Tagetes erecta] AAM45381 21360357 beta cyclase [Tagetes erecta] AAM14335 20259239 putative lycopene beta cyclase [Arabidopsis thaliana] AAO18661 27728515 lycopene beta-cyclase [Zea mays] AAA81880 735882 lycopene cyclase Q43503 11133022 Lycopene beta cyclase, chloroplast precursor S66350 2129931 lycopene beta-cyclase (EC 5.5.1.—) - tomato XP_464409 50905841 putative lycopene beta-cyclase [Oryza sativa (japonica cultivar-group)] CAD70565 45237491 lycopene cyclase [Bixa orellana] Q43578 11133025 Lycopene beta cyclase, chloroplast precursor AAL92175 19569782 beta-lycopene cyclase [Sandersonia aurantiaca] AAX54906 61742130 putative chloroplast lycopene beta cyclase precursor [Chlamydomonas reinhardtii] S66349 2129954 lycopene beta-cyclase (EC 5.5.1.—) - common tobacco AAG21133 10644119 chromoplast-specific lycopene beta-cyclase [Lycopersicon esculentum] CAB92977 8247354 neoxanthin synthase [Solanum tuberosum] CAB93342 8249885 neoxanthin synthase [Lycopersicon esculentum] Q9SEA0 11131528 Capsanthin/capsorubin synthase, chloroplast precursor Q42435 12643508 Capsanthin/capsorubin synthase, chloroplast precursor AAO64977 37730608 lycopene beta cyclase [Haematococcus pluvialis] Q40424 11133011 Lycopene beta cyclase, chloroplast precursor ABB52072 79154940 putative capsanthin-capsorubin synthase [Daucus carota subsp. sativus] AAQ02668 33304511 lycopene cyclase [Setaria italica] CAA54961 840729 putative chromoplastic oxydo-reductase [Capsicum annuum] EAJ62838 44489136 unknown [environmental sequence] YP_401079 81300871 Lycopene cyclase, beta and epsilon [Synechococcus elongatus PCC 7942] YP_172741 56752040 lycopene cyclase [Synechococcus elongatus PCC 6301] ZP_011 . . . 88808972 lycopene beta cyclase [Synechococcus sp. WH 7805] EAK50052 44615956 unknown [environmental sequence] NP_892751 33861190 putative lycopene epsilon cyclase [Prochlorococcus marinus subsp. pastoris str. CCMP1986] NP_875182 33240240 Lycopene epsilon cyclase [Prochlorococcus marinus subsp. marinus str. CCMP1375] YP_382237 78213458 Lycopene cyclase, beta and epsilon [Synechococcus sp. CC9605] YP_397130 78779018 Lycopene cyclase, beta and epsilon [Prochlorococcus marinus str. MIT 9312] NP_896821 33865262 lycopene beta cyclase [Synechococcus sp. WH 8102] YP_397570 78779458 Lycopene cyclase, beta and epsilon [Prochlorococcus marinus str. MIT 9312] ZP_010 . . . 87302144 lycopene cyclase [Synechococcus sp. WH 5701] EAK17149 44569190 unknown [environmental sequence] YP_291882 72382527 lycopene cyclase, beta and epsilon [Prochlorococcus marinus str. NATL2A] NP_875528 33240586 Lycopene beta cyclase related dehydrogenase [Prochlorococcus marinus subsp. marinus str. CCMP1375] NP_893181 33861620 putative lycopene beta cyclase [Prochlorococcus marinus subsp. pastoris str. CCMP1986] NP_895600 33864040 putative lycopene epsilon cyclase [Prochlorococcus marinus str. MIT 9313] EAI47456 44325573 unknown [environmental sequence] YP_291268 72381913 lycopene cyclase, beta and epsilon [Prochlorococcus marinus str. NATL2A] ZP_010 . . . 84517806 Lycopene beta cyclase related dehydrogenase [Prochlorococcus marinus str. MIT 9211] AAF34191 6970079 lycopene epsilon cyclase [Daucus carota] ZP_010 . . . 84518202 Lycopene epsilon cyclase [Prochlorococcus marinus str. MIT 9211] YP_376736 78184301 Lycopene cyclase, beta and epsilon [Synechococcus sp. CC9902] ZP_003 . . . 66796756 Lycopene cyclase, beta and epsilon [Deinococcus geothermalis DSM 11300] NP_894954 33863394 putative lycopene beta cyclase [Prochlorococcus marinus str. MIT 9313] AAT76051 50365502 lycopene cyclase [Citrus clementina] EAK22047 44576122 unknown [environmental sequence] NP_294525 15805827 lycopene cyclase [Deinococcus radiodurans R1]

TABLE 24 Examples of carotenoid glucosyltransferase polypeptides. ACCESSION GI PROTEIN DESCRIPTION AAA21261 148395 CrtX [Pantoea agglomerans] AAN85597 27228291 Zeaxanthin Glucosyl Transferase [Pantoea stewartii] BAB79601 18143446 crtX [Pantoea agglomerans pv. milletiae] AAZ73147 72536082 zeaxanthin glucosyl transferase [Enterobacteriaceae bacterium DC413] AAZ73128 72536060 zeaxanthin glucosyl transferase [Enterobacteriaceae bacterium DC260] AAZ73140 72536074 zeaxanthin glucosyl transferase [Enterobacteriaceae bacterium DC416] Q01330 231911 Zeaxanthin glucosyl transferase ZP_006 . . . 71674312 UDP-glycosyltransferase, MGT [Trichodesmium erythraeum IMS101] NP_439972 16329244 zeaxanthin glucosyl transferase [Synechocystis sp. PCC 6803] EAH29368 44130903 unknown [environmental sequence] ZP_005 . . . 67926135 zeaxanthin glucosyl transferase, hypothetical protein [Crocosphaera watsonii WH 8501] YP_378763 78188425 hypothetical protein Cag_0447 [Chlorobium chlorochromatii CaD3] ZP_005 . . . 68549418 Glycosyl transferase, group 1 [Pelodictyon phaeoclathratiforme BU-1] ZP_010 . . . 85713606 glycosyl transferase, group 1 [Nitrobacter sp. Nb-311A] YP_317171 75674750 glycosyl transferase, group 1 [Nitrobacter winogradskyi Nb- 255] ZP_006 . . . 69929171 Glycosyl transferase, group 1 [Nitrobacter hamburgensis X14] ZP_009 . . . 84500589 hypothetical protein OB2597_11541 [Oceanicola batsensis HTCC2597] ZP_009 . . . 83953176 hypothetical protein NAS141_12746 [Sulfitobacter sp. NAS- 14.1] ZP_009 . . . 83942121 hypothetical protein EE36_07793 [Sulfitobacter sp. EE-36] YP_508020 89052569 glycosyl transferase, group 1 [Jannaschia sp. CCS1] ZP_010 . . . 85704103 hypothetical protein ROS217_13931 [Roseovarius sp. 217] ZP_009 . . . 83370850 probable glycosyltransferase [Rhodobacter sphaeroides ATCC 17025] ZP_006 . . . 69934465 Glycosyl transferase, group 1 [Paracoccus denitrificans PD1222] ZP_009 . . . 83949880 probable glycosyltransferase [Roseovarius nubinhibens ISM] YP_376237 78183803 putative glycosyltransferase [Synechococcus sp. CC9902] YP_376129 78183695 probable glycosyltransferase [Synechococcus sp. CC9902] YP_374296 78186253 hypothetical protein Plut_0365 [Pelodictyon luteolum DSM 273] ZP_010 . . . 87301651 Putative glycosyltransferase [Synechococcus sp. WH 5701] ZP_011 . . . 88809938 Putative glycosyltransferase [Synechococcus sp. WH 7805] BAE47471 78483937 carotenoid glucosyltransferase [Paracoccus sp. N81106] ZP_010 . . . 87303273 probable glycosyltransferase [Synechococcus sp. WH 5701] YP_376127 78183693 probable glycosyltransferase [Synechococcus sp. CC9902] YP_501334 88196509 hypothetical protein SAOUHSC_02880 [Staphylococcus aureus subsp. aureus NCTC 8325] YP_187370 57652300 glycosyl transferase, group 2 family protein [Staphylococcus aureus subsp. aureus COL] CAA66627 1340131u nnamed protein product [Staphylococcus aureus] YP_041987 49484763 putative glycosyl transferase [Staphylococcus aureus subsp. aureus MRSA252] YP_417885 82752144 hypothetical protein SAB2436c [Staphylococcus aureus RF122] YP_252404 70725490 hypothetical protein SH0489 [Staphylococcus haemolyticus JCSC1435] NP_693379 23099913 hypothetical protein OB2458 [Oceanobacillus iheyensis HTE831] ZP_008 . . . 82501285 conserved hypothetical protein [Caldicellulosiruptor saccharolyticus DSM 8903] ZP_010 . . . 87303565 hypothetical protein WH5701_09900 [Synechococcus sp. WH 5701]

TABLE 25 Examples of acyl CoA:diacyglycerol acyltransferase (DGAT) polypeptides. ACCESSION GI PROTEIN DESCRIPTION XP_957022 85082953 hypothetical protein [Neurospora crassa N150] XP_386864 46124621 hypothetical protein FG06688.1 [Gibberella zeae PH-1] XP_755172 71000982 diacylglycerol O-acyltransferase DGAT [Aspergillus fumigatus Af293] XP_663763 67539978 hypothetical protein AN6159.2 [Aspergillus nidulans FGSC A4] BAE65302 83775179 unnamed protein product [Aspergillus oryzae] XP_502557 50550169 hypothetical protein [Yarrowia lipolytica] AAS78662 56199782 diacylglycerol acyltransferase [Glycine max] ABB84383 82582915 diacylglycerol acyltransferase [Jatropha curcas] AAV31083 54145459 1,2-diacyl-sn-glycerol:acyl-CoA acyltransferase [Euonymus alatus] AAG23696 10803053 diacylglycerol acyltransferase [Perilla frutescens] AAF64065 7576941 putative diacylglycerol acyltransferase [Brassica napus] AAS01606 41387497 acyl-CoA:diacylglycerol acyltransferase 1 [Olea europaea] AAT73629 50299542 acyl CoA:diacylglycerol acyltransferase [Glycine max] AAM03340 67043496 putative diacylglycerol acyltransferase [Tropaeolum majus] XP_645633 66824557 hypothetical protein DDB0202877 [Dictyostelium discoideum] AAF19345 6625653 diacylglycerol acylCoA acyltransferase [Nicotiana tabacum] AAY40785 63376239 diacylglycerol acyltransferase DGAT2 [Brassica juncea] AAW47581 57231736 diacylglycerol acyltransferase [Oryza sativa (japonica cultivar-group)] AAR11479 38146080 diacylglycerol acyltransferase [Ricinus communis] AAY40784 63376226 diacylglycerol acyltransferase DGAT1 [Brassica juncea] AAP68322 31711932 At2g19450 [Arabidopsis thaliana] AAW51456 57545061 diacylglycerol acyltransferase [Lotus corniculatus var. japonicus] AAD45536 5579408 putative diacylglycerol acyltransferase [Brassica napus] BAD53762 53791817 putative acyl-CoA:diacylglycerol acyltransferase [Oryza sativa (japonica cultivar-group)] NP_956024 41054343 hypothetical protein LOC325875 [Danio rerio] AAL49962 18642598 diacylglycerol acyltransferase 1 [Bos taurus] XP_930884 89028385 similar to Diacylglycerol O-acyltransferase 1 (Diglyceride acyltransferase) (ACAT-related gene) [Homo sapiens] NP_777118 27819636 diacylglycerol O-acyltransferase 1 [Bos taurus] Q9GMF1 18202926 Diacylglycerol O-acyltransferase 1 (Diglyceride acyltransferase) NP_036211 6912332 diacylglycerol O-acyltransferase 1 [Homo sapiens] AAH06263 34782946 DGAT1 protein [Homo sapiens] XP_780515 72006039 similar to Diacylglycerol O-acyltransferase 1 [Strongylocentrotus purpuratus] AAD40881 5225382 putative diacylglycerol acyltransferase [Brassica napus] XP_539214 73974769 similar to Diacylglycerol O-acyltransferase 1 (ACAT related gene product 1) isoform 1 [Canis familiaris] AAZ22403 71063860 diacylglycerol O-acyltransferase 1 [Bubalus bubalis] NP_999216 47522918 diacylglycerol acyltransferase [Sus scrofa] NP_001 . . . 50539976 hypothetical protein LOC436731 [Danio rerio] XP_849176 73974767 similar to Diacylglycerol O-acyltransferase 1 (ACAT related gene product 1) isoform 2 [Canis familiaris] NP_505828 71997360 H19N07.4 [Caenorhabditis elegans] AAF82410 9049538 diacylglycerol acyltransferase [Caenorhabditis elegans] CAE75170 39591950 Hypothetical protein CBG23107 [Caenorhabditis briggsae] XP_626337 66358318 diacylglycerol acyltransferase 1 [Cryptosporidium parvum Iowa II] XP_668402 67624239 acyl-CoA:diacylglycerol acyltransferase 1-related enzyme [Cryptosporidium hominis TU502] AAP94208 33113253 acyl-CoA:diacylglycerol acyltransferase 1-related enzyme [Toxoplasma gondii] AAP94209 33113255 acyl-CoA:diacylglycerol acyltransferase 1-related enzyme [Toxoplasma gondii] XP_579557 62652535 PREDICTED: diacylglycerol O-acyltransferase 1 [Rattus norvegicus] BAC66171 29170489 diacylglycerol acyltransferase [Mus musculus] Q9ERM3 18202872 Diacylglycerol O-acyltransferase 1 (Diglyceride acyltransferase) AAL78366 18698659 acyl coenzyme A:diacylglycerol acyltransferase [Drosophila melanogaster] NP_995724 45552403 CG31991-PD, isoform D [Drosophila melanogaster] NP_724017 24584734 CG31991-PC, isoform C [Drosophila melanogaster] XP_858062 73974765 similar to Diacylglycerol O-acyltransferase 1 (ACAT related gene product 1) isoform 3 [Canis familiaris] XP_728984 82915156 hypothetical protein PY01256 [Plasmodium yoelii yoelii str. 17XNL] CAG11944 47225461 unnamed protein product [Tetraodon nigroviridis] BAD27526 50199438 acyl-CoA:diacylglycerol acyltransferase [eukaryotic synthetic construct] XP_317656 31226099 ENSANGP00000002281 [Anopheles gambiae str. PEST] AAV59457 55733950 putative diacylglycerol acyltransferase [Oryza sativa (japonica cultivar-group)] EAL33593 54644853 GA16599-PA [Drosophila pseudoobscura] XP_678753 68073677 diacylglycerol O-acyltransferase [Plasmodium berghei strain ANKA] XP_520014 55631434 PREDICTED: similar to Diacylglycerol O-acyltransferase 1 (Diglyceride acyltransferase) [Pan troglodytes] CAG10815 47219451 unnamed protein product [Tetraodon nigroviridis] XP_624754 66522700 PREDICTED: similar to ENSANGP00000002281 [Apis mellifera] CAC69884 15620769 diacylglycerol acyltransferase I [Rattus norvegicus] XP_686181 68363630 PREDICTED: similar to Diacylglycerol O-acyltransferase 1 (Diglyceride acyltransferase) [Danio rerio] XP_734008 70921323 diacylglycerol O-acyltransferase [Plasmodium chabaudi chabaudi] XP_673128 68062248 hypothetical protein PB300300.00.0 [Plasmodium berghei strain ANKA] AAS72376 45642963 acyl-CoA:cholesterol acyltransferase beta [Toxoplasma gondii] AAS72375 45642961 acyl-CoA:cholesterol acyltransferase alpha [Toxoplasma gondii] NP_586145 19074639 STEROL O-ACYLTRANSFERASE [Encephalitozoon cuniculi GB-M1] XP_640280 66812202 hypothetical protein DDB0205259 [Dictyostelium discoideum] AAY40783 63376221 diacylglycerol acyltransferase [Brassica juncea] XP_765774 71032265 diacylglycerol O-acyltransferase [Theileria parva strain Muguga] Q876L2 34582301 Sterol O-acyltransferase 2 (Sterol-ester synthase 2) XP_571260 58268208 sterol O-acyltransferase [Cryptococcus neoformans var. neoformans JEC21] EAL20032 50257323 hypothetical protein CNBF3580 [Cryptococcus neoformans var. neoformans B-3501A] XP_954478 84999514 acyl transferase [Theileria annulata strain Ankara] XP_505086 50555355 hypothetical protein [Yarrowia lipolytica] NP_588558 19076058 hypothetical protein SPCP1E11.05c [Schizosaccharomyces pombe 972h-] AAC49441 1389739 acyl-CoA:sterol acyltransferase NP_014416 6324346 Acyl-CoA:sterol acyltransferase, isozyme of Are1p; Are2p [Saccharomyces cerevisiae] XP_750354 70991010 sterol o-acyltransferase APE2 [Aspergillus fumigatus Af293] XP_382192 46110268 hypothetical protein FG02016.1 [Gibberella zeae PH-1] BAE54934 83764790 unnamed protein product [Aspergillus oryzae] XP_885914 76617939 similar to Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (ACAT-2) isoform 2 [Bos taurus] XP_591251 76617937 similar to Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (ACAT-2) isoform 1 [Bos taurus] BAC00846 21392392 AcylCoA:Cholesterol Acyltransferase 2 [Rattus norvegicus] NP_649816 28571583 CG8112-PA [Drosophila melanogaster] NP_666176 22122547 sterol O-acyltransferase 2 [Mus musculus] O88908 18202245 Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (ACAT-2) XP_761502 71022545 hypothetical protein UM05355.1 [Ustilago maydis 521] NP_714950 40254723 sterol O-acyltransferase 2 [Rattus norvegicus] EAQ86094 88178626 hypothetical protein CHGG_07347 [Chaetomium globosum CBS 148.51] XP_461395 50425599 hypothetical protein DEHA0F25652g [Debaryomyces hansenii CBS767] XP_661812 67527926 hypothetical protein AN4208.2 [Aspergillus nidulans FGSC A4] AAH96091 64654094 Sterol O-acyltransferase 2 [Homo sapiens] O75908 18202149 Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (ACAT-2) AAH96090 64652990 Sterol O-acyltransferase 2 [Homo sapiens] AAK48829 13898623 acyl coenzyme A: cholesterol acyltransferase-2 [Homo sapiens] XP_543637 73996435 PREDICTED: similar to sterol O-acyltransferase 2 [Canis familiaris] O77759 18202176 Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (ACAT-2) AAO32474 28564191 ARE2 [Saccharomyces castellii] XP_323485 32405744 hypothetical protein [Neurospora crassa] NP_982606 45184888 AAR065Cp [Eremothecium gossypii] NP_593708 19114620 hypothetical protein SPAC13G7.06 [Schizosaccharomyces pombe 972h-] AAO32554 28564940 ARE2 [Saccharomyces kluyveri] EAL28962 54639560 GA20833-PA [Drosophila pseudoobscura] XP_449806 50294790 hypothetical protein CAGL0M10571g [Candida glabrata CBS138] NP_033256 84619697 sterol O-acyltransferase 1 [Mus musculus] Q61263 18202591 Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (ACAT-1) BAC34925 26342537 unnamed protein product [Mus musculus] XP_452607 50305295 unnamed protein product [Kluyveromyces lactis] NP_001 . . . 77735363 hypothetical protein LOC504287 [Bos taurus] Q60457 18202585 Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (ACAT-1) XP_320321 58393811 ENSANGP00000016512 [Anopheles gambiae str. PEST] XP_320320 58393809 ENSANGP00000016486 [Anopheles gambiae str. PEST] O70536 18202126 Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (ACAT-1) XP_714776 68482533 acyl-CoA cholesterol acyltransferase [Candida albicans SC5314] P84285 56404462 Sterol O-acyltransferase 2 (Sterol-ester synthase) (ASAT) AAH77916 50416229 Soat1-prov protein [Xenopus laevis] XP_692855 68364838 PREDICTED: similar to Soat1-prov protein [Danio rerio] CAI13574 55960156 sterol O-acyltransferase (acyl-Coenzyme A: cholesterol acyltransferase) 1 [Homo sapiens] AAL56227 18028942 cholesterol acyltransferase 1 [Gorilla gorilla] AAL56228 18028944 cholesterol acyltransferase 1 [Pongo pygmaeus] AAC37532 4878022 acyl-coenzyme A: cholesterol acyltransferase [Homo sapiens] 2201440A 1585676 acyl-CoA/cholesterol acyltransferase Q876L3 34582302 Sterol O-acyltransferase 1 (Sterol-ester synthase 1) BAE01048 67969393 unnamed protein product [Macaca fascicularis] XP_514030 55588858 PREDICTED: hypothetical protein XP_514030 [Pan troglodytes] XP_547445 73961286 similar to Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (ACAT-1) [Canis familiaris] EAQ84619 88177151 hypothetical protein CHGG_08633 [Chaetomium globosum CBS 148.51] O77761 18202178 Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (ACAT-1) XP_422267 50751122 PREDICTED: similar to Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (ACAT-1) [Gallus gallus] XP_693284 68392980 PREDICTED: similar to Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (ACAT-1) [Danio rerio] AAT92940 51013293 YCR048W [Saccharomyces cerevisiae] XP_956576 85080625 hypothetical protein [Neurospora crassa N150] XP_624691 66564061 PREDICTED: similar to ENSANGP00000016486 [Apis mellifera] CAF96514 47222847 unnamed protein product [Tetraodon nigroviridis] XP_788209 72085563 PREDICTED: similar to sterol O-acyltransferase 1 [Strongylocentrotus purpuratus] XP_445307 50285757 unnamed protein product [Candida glabrata] CAE70002 39596364 Hypothetical protein CBG16409 [Caenorhabditis briggsae] CAG07990 47225647 unnamed protein product [Tetraodon nigroviridis] NP_510623 17549960 B0395.2 [Caenorhabditis elegans] AAX28331 76157393 SJCHGC04421 protein [Schistosoma japonicum] CAI96158 66347204 Diacylglycerol O-acyltransferase [Bubalus bubalis] XP_390039 46136695 hypothetical protein FG09863.1 [Gibberella zeae PH-1] XP_643169 66819019 hypothetical protein DDB0203882 [Dictyostelium discoideum] AAO53095 28850306 hypothetical protein [Dictyostelium discoideum] AAB06959 1515472 acyl-CoA:cholesterol acyltransferase [Oryctolagus cuniculus] NP_945619 39933343 putative alginate o-acetyltransferase AlgI [Rhodopseudomonas palustris CGA009] ZP_008 . . . 77691302 Membrane bound O-acyl transferase, MBOAT [Rhodopseudomonas palustris BisB5] XP_465546 50908115 putative wax synthase [Oryza sativa (japonica cultivar- group)]

TABLE 29 Examples of Prenyldiphosphate synthase polypeptides Accession GI Description 29A: Bacteria Proteins that require a mitochondrial targeting sequence ZP_009 . . . 83373595 Trans-hexaprenyltranstransferase [Rhodobacter sphaeroides ATCC 17029] ZP_009 . . . 83371280 Trans-hexaprenyltranstransferase [Rhodobacter sphaeroides ATCC 17025] CAD24417 20429105 decaprenyl diphosphate synthase [Paracoccus zeaxanthinifaciens] ZP_010 . . . 85705714 Geranylgeranyl pyrophosphate synthase/Polyprenyl synthetase [Roseovarius sp. 217] ZP_010 . . . 84515724 decaprenyl diphosphate synthase [Loktanella vestfoldensis SKA53] YP_165582 56695234 decaprenyl diphosphate synthase [Silicibacter pomeroyi DSS-3] ZP_010 . . . 86139019 decaprenyl diphosphate synthase [Roseobacter sp. MED193] ZP_009 . . . 83941379 decaprenyl diphosphate synthase [Sulfitobacter sp. EE-36] ZP_009 . . . 83854856 decaprenyl diphosphate synthase [Sulfitobacter sp. NAS-14.1] ZP_006 . . . 69299873 Farnesyltranstransferase [Silicibacter sp. TM1040] ZP_010 . . . 84683979 Geranylgeranyl pyrophosphate synthase/Polyprenyl synthetase [Rhodobacterales bacterium HTCC2654] ZP_009 . . . 84500217 decaprenyl diphosphate synthase [Oceanicola batsensis HTCC2597] ZP_009 . . . 83952381 decaprenyl diphosphate synthase [Roseovarius nubinhibens ISM] ZP_006 . . . 69937106 Trans-hexaprenyltranstransferase [Paracoccus denitrificans PD1222] ZP_005 . . . 68180845 Trans-hexaprenyltranstransferase [Jannaschia sp. CCS1] ZP_008 . . . 78495595 Polyprenyl synthetase [Rhodopseudomonas palustris BisB18] AAY82368 67866738 decaprenyl diphosphate synthase [Agrobacterium tumefaciens] NP_353656 15887975 hypothetical protein AGR_C_1125 [Agrobacterium tumefaciens str. C58] ZP_008 . . . 77688465 Farnesyltranstransferase [Rhodopseudomonas palustris BisB5] NP_531334 17934544 octaprenyl-diphosphate synthase [Agrobacterium tumefaciens str. C58] YP_484709 86748213 Farnesyltranstransferase [Rhodopseudomonas palustris HaA2] AAP56240 37903500 decaprenyl diphosphate synthase [Agrobacterium tumefaciens] YP_192388 58040424 Decaprenyl diphosphate synthase [Gluconobacter oxydans 621H] 29B: Subunit 1- Proteins that contain mitochondrial targeting sequence T43193 11279237 trans-pentaprenyltranstransferase homolog - fission yeast (Schizosaccharomyces pombe ) AAD28559 4732024 trans-prenyltransferase [Homo sapiens] AAI07275 78070698 Trans-prenyltransferase [Mus musculus] BAE48216 81157931 subunit 1 of decaprenyl diphosphate synthase [Homo sapiens] AAH49211 29165656 PDSS1 protein [Homo sapiens] Q33DR2 85700953 Decaprenyl-diphosphate synthase subunit 1 (Solanesyl-diphosphate synthase subunit 1) (Trans-prenyltransferase) XP_507706 55633583 PREDICTED: similar to TPRT protein [Pan troglodytes] XP_586717 76632198 PREDICTED: similar to trans-prenyltransferase [Bos taurus] XP_849908 73948851 PREDICTED: similar to trans-prenyltransferase [Canis familiaris] 29C: Subunit 2- Proteins that contain mitochondrial targeting sequence O13851 60389474 Decaprenyl-diphosphate synthase subunit 2 (Decaprenyl pyrophosphate synthetase subunit 2) BAE48218 81157935 subunit 2 of solanesyl diphosphate synthase [Mus musculus] BAE48217 81157933 subunit 2 of decaprenyl diphosphate synthase [Homo sapiens]

TABLE 30 Examples of PHB-Polyprenyltransferase polypeptides GI PROTEIN DESCRIPTION 51013645 YNR041C [Saccharomyces cerevisiae] 50285815 unnamed protein product [Candida glabrata] 50311051 unnamed protein product [Kluyveromyces lactis] 45200866 AGL231Wp [Eremothecium gossypii] 50555263 hypothetical protein [Yarrowia lipolytica] 68473193 para-hydroxybenzoate: polyprenyl transferase [Candida albicans SC5314] 50410039 hypothetical protein DEHA0A14212g [Debaryomyces hansenii CBS767] 83769349 unnamed protein product [Aspergillus oryzae] 70994900 para-hydroxybenzoate-polyprenyltransferase precursor [Aspergillus fumigatus Af293] 19114131 hypothetical protein SPAC56F8. 04c [Schizosaccharomyces pombe 972h-] 39973573 hypothetical protein MG01067. 4 [Magnaporthe grisea 70-15] 85078920 protein related to para-hydroxybenzoate polyprenyltransferase precursor [Neurospora crassa N150] 76660839 PREDICTED: similar to para-hydroxybenzoate-polyprenyltransferase, mitochondrial [Bos taurus] 52138578 para-hydroxybenzoate-polyprenyltransferase, mitochondrial [Homo sapiens] 18088424 COQ2 protein [Homo sapiens] 47221448 unnamed protein product [Tetraodon nigroviridis] 58385249 ENSANGP00000012220 [Anopheles gambiae str. PEST] 50746583 PREDICTED: similar to hypothetical protein CL640 [Gallus gallus] 54638587 GA21912-PA [Drosophila pseudoobscura] 21355567 CG9613-PA [Drosophila melanogaster] 71005862 hypothetical protein UM01450. 1 [Ustilago maydis 521]

TABLE 31 Examples of soluble transydrogenase polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAD40691 5163507 soluble pyridine nucleotide transhydrogenase [Azotobacter vinelandii 2 ZP_013 . . . 107102541 hypothetical protein PaerPA_01003605 [Pseudomonas aeruginosa PACS2] 3 ZP_012 . . . 94416745 hypothetical protein PaerP_01001583 [Pseudomonas aeruginosa PA7] 4 O05139 11135075 Soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 5 YP_259077 70729340 soluble pyridine nucleotide transhydrogenase [Pseudomonas fluorescens Pf-5] 6 YP_349590 77460083 soluble pyridine nucleotide transhydrogenase [Pseudomonas fluorescens PfO-1] 7 YP_274087 71735155 soluble pyridine nucleotide transhydrogenase [Pseudomonas syringae pv. phaseolicola 1448A] 8 YP_234983 66045142 soluble pyridine nucleotide transhydrogenase [Pseudomonas syringae pv. syringae B728a] 9 YP_609219 104782721 soluble pyridine nucleotide transhydrogenase (NAD(P)(+) transhydrogenase [B-specific]) [Pseudomonas entomophila L48] 10 Q884I6 38258471 Soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 11 ZP_009 . . . 82739140 soluble pyridine nucleotide transhydrogenase [Pseudomonas putida F1] 12 YP_573629 92113701 pyridine nucleotide-disulphide oxidoreductase dimerisation region [Chromohalobacter salexigens DSM 3043] 13 ZP_008 . . . 77951906 NAD(P) transhydrogenase [Marinobacter aquaeolei VT8] 14 YP_433907 83645472 Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzyme [Hahella chejuensis KCTC 2396] 15 ZP_011 . . . 89094310 soluble pyridine nucleotide transhydrogenase [Oceanospirillum sp. MED92] 16 ZP_013 . . . 94499566 Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzyme [Oceanobacter sp. RED65] 17 ZP_011 . . . 88800089 soluble pyridine nucleotide transhydrogenase [Reinekea sp. MED297] 18 ZP_010 . . . 87121312 soluble pyridine nucleotide transhydrogenase [Marinomonas sp. MED121] 19 ZP_012 . . . 90417525 soluble pyridine nucleotide transhydrogenase [marine gamma proteobacterium HTCC2207] 20 Q7MQ83 47606104 Soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 21 YP_527277 90021450 regulatory protein, ArsR [Saccharophagus degradans 2-40] 22 ZP_009 . . . 83586194 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Escherichia coli 101-1] 23 YP_219002 62182585 soluble pyridine nucleotide transhydrogenase [Salmonella enterica subsp. enterica serovar Choleraesuis str. SC-B67] 24 YP_543498 91213512 soluble pyridine nucleotide transhydrogenase [Escherichia coli UTI89] 25 Q87KN5 33301640 Soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 26 YP_153038 56415963 soluble pyridine nucleotide transhydrogenase [Salmonella enterica subsp. enterica serovar Paratyphi A str. ATCC 9150] 27 YP_672034 110644304 soluble pyridine nucleotide transhydrogenase [Escherichia coli 536] 28 ZP_013 . . . 106883834 FAD-dependent pyridine nucleotide-disulphide oxidoreductase:Pyridine nucleotide-disulphide oxidoreductase dimerisation region:FAD dependent oxidoreductase [Psychromonas ingrahamii 37] 29 ZP_006 . . . 75176647 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Shigella boydii BS512] 30 Q8FB93 38258590 Soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 31 ZP_007 . . . 75197835 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Escherichia coli HS] 32 ZP_007 . . . 75513852 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Escherichia coli 53638] 33 Q8X727 21362954 Soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 34 YP_267100 71278420 soluble pyridine nucleotide transhydrogenase [Colwellia psychrerythraea 34H] 35 Q83MI1 39932373 Soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 36 ZP_007 . . . 75238410 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Escherichia coli E110019] 37 ZP_007 . . . 75854141 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Vibrio sp. Ex25] 38 YP_052329 50123162 soluble pyridine nucleotide transhydrogenase [Erwinia carotovora subsp. atroseptica SCRI1043] 39 ZP_008 . . . 77973002 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Yersinia frederiksenii ATCC 33641] 40 ZP_008 . . . 77957412 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Yersinia bercovieri ATCC 43970] 41 ZP_008 . . . 78702791 NAD(P) transhydrogenase (B-specific) [Alkalilimnicola ehrlichei MLHE-1] 42 YP_645986 108810219 soluble pyridine nucleotide transhydrogenase [Yersinia pestis Nepal516] 43 ZP_007 . . . 75828016 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Vibrio cholerae O395] 44 Q7MBG9 47606102 Soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 45 ZP_008 . . . 77976809 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Yersinia intermedia ATCC 29909] 46 ZP_008 . . . 77961066 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Yersinia mollaretii ATCC 43969] 47 ZP_010 . . . 86148649 soluble pyridine nucleotide transhydrogenase [Vibrio sp. MED222] 48 YP_205822 59713046 soluble pyridine nucleotide transhydrogenase [Vibrio fischeri ES114] 49 ZP_009 . . . 84394322 pyridine nucleotide-disulfide oxidoreductase, class I [Vibrio splendidus 12B01] 50 ZP_011 . . . 89074500 soluble pyridine nucleotide transhydrogenase [Photobacterium sp. SKA34] 51 YP_154714 56459433 soluble pyridine nucleotide transhydrogenase [Idiomarina loihiensis L2TR] 52 YP_046885 50085375 soluble pyridine nucleotide transhydrogenase [Acinetobacter sp. ADP1] 53 ZP_012 . . . 90581098 soluble pyridine nucleotide transhydrogenase [Vibrio angustum S14] 54 YP_693290 110834431 NAD(P) transhydrogenase [Alcanivorax borkumensis SK2] 55 ZP_011 . . . 88795508 soluble pyridine nucleotide transhydrogenase [Alteromonas macleodii ‘Deep ecotype’] 56 ZP_012 . . . 90407259 soluble pyridine nucleotide transhydrogenase [Psychromonas sp. CNPT3] 57 E65203 7431869 probable dehydrogenase (EC 1.8.1.—) udhA - Escherichia coli (strain K-12) 58 YP_410260 82546313 putative oxidoreductase [Shigella boydii Sb227] 59 YP_312883 74314464 putative oxidoreductase [Shigella sonnei Ss046] 60 YP_131541 54310521 soluble pyridine nucleotide transhydrogenase [Photobacterium profundum SS9] 61 AAC43068 396309 ORF_f444 [Escherichia coli] 62 H86087 25284914 probable oxidoreductase udhA [imported] - Escherichia coli (strain O157:H7, substrain EDL933) 63 ZP_012 . . . 90414698 soluble pyridine nucleotide transhydrogenase [Photobacterium profundum 3TCK] 64 NP_709766 56480495 soluble pyridine nucleotide transhydrogenase [Shigella flexneri 2a str. 301] 65 YP_405233 82778884 putative oxidoreductase [Shigella dysenteriae Sd197] 66 YP_691341 110807821 putative oxidoreductase [Shigella flexneri 5 str. 8401] 67 NP_667661 22124238 soluble pyridine nucleotide transhydrogenase [Yersinia pestis KIM] 68 YP_341375 77361800 soluble pyridine nucleotide transhydrogenase [Pseudoalteromonas haloplanktis TAC125] 69 YP_455837 85060135 soluble pyridine nucleotide transhydrogenase [Sodalis glossinidius str. ‘morsitans’] 70 ZP_012 . . . 91771448 FAD-dependent pyridine nucleotide-disulphide oxidoreductase:Pyridine nucleotide-disulphide oxidoreductase dimerisation region:FAD dependent oxidoreductase [Psychrobacter sp. PRwf-1] 71 ZP_011 . . . 88861405 soluble pyridine nucleotide transhydrogenase [Pseudoalteromonas tunicata D2] 72 YP_264617 71065890 soluble pyridine nucleotide transhydrogenase [Psychrobacter arcticus 273-4] 73 YP_580301 93005864 pyridine nucleotide-disulphide oxidoreductase dimerisation region [Psychrobacter cryohalolentis K5] 74 ZP_010 . . . 85710769 soluble pyridine nucleotide transhydrogenase [Idiomarina baltica OS145] 75 ZP_012 . . . 91227486 soluble pyridine nucleotide transhydrogenase [Vibrio alginolyticus 12G01] 76 ZP_007 . . . 75814072 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Vibrio cholerae V52] 77 YP_666832 110670275 soluble pyridine nucleotide transhydrogenase [Francisella tularensis subsp. tularensis FSC 198] 78 YP_169700 56707804 soluble pyridine nucleotide transhydrogenase [Francisella tularensis subsp. tularensis SCHU S4] 79 YP_513671 89256309 soluble pyridine nucleotide transhydrogenase [Francisella tularensis subsp. holarctica] 80 CAA46822 43237 udhA [Escherichia coli] 81 ZP_005 . . . 67929855 FAD-dependent pyridine nucleotide-disulphide oxidoreductase:Pyridine nucleotide-disulphide oxidoreductase dimerisation region [Solibacter usitatus Ellin6076] 82 YP_592298 94970250 pyridine nucleotide-disulphide oxidoreductase [Acidobacteria bacterium Ellin345] 83 NP_961763 41408927 soluble pyridine nucleotide transhydrogenase [Mycobacterium avium subsp. paratuberculosis K-10] 84 P66006 54042093 Probable soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 85 ZP_007 . . . 76783189 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Mycobacterium tuberculosis F11] 86 YP_716291 111225497 soluble pyridine nucleotide transhydrogenase [Frankia alni ACN14a] 87 ZP_005 . . . 68235154 FAD-dependent pyridine nucleotide-disulphide oxidoreductase:Pyridine nucleotide-disulphide oxidoreductase dimerisation domain [Frankia sp. EAN1pec] 88 ZP_012 . . . 90205475 FAD-dependent pyridine nucleotide-disulphide oxidoreductase:Pyridine nucleotide-disulphide oxidoreductase dimerisation region [Mycobacterium vanbaalenii PYR-1] 89 YP_482962 86742562 pyridine nucleotide-disulphide oxidoreductase dimerisation region [Frankia sp. CcI3] 90 NP_532346 17935556 soluble pyridine nucleotide transhydrogenase [Agrobacterium tumefaciens str. C58] 91 AAY87873 68165826 pyridine nucleotide-disulfide oxidoreductase class I [Vibrio cholerae] 92 AAY87863 68165806 pyridine nucleotide-disulfide oxidoreductase class I [Vibrio cholerae] 93 AAY87865 68165810 pyridine nucleotide-disulfide oxidoreductase class I [Vibrio cholerae] 94 YP_469365 86357473 probable pyridine nucleotide transhydrogenase protein [Rhizobium etli CFN 42] 95 ZP_010 . . . 85706062 soluble pyridine nucleotide transhydrogenase [Roseovarius sp. 217] 96 ZP_014 . . . 113873124 putative soluble pyridine nucleotide transhydrogenase [Sinorhizobium medicae WSM419] 97 NP_385835 15965482 soluble pyridine nucleotide transhydrogenase [Sinorhizobium meliloti 1021] 98 YP_169016 56698639 soluble pyridine nucleotide transhydrogenase [Silicibacter pomeroyi DSS-3] 99 ZP_009 . . . 83951172 soluble pyridine nucleotide transhydrogenase [Roseovarius nubinhibens ISM] 100 YP_611681 99078423 pyridine nucleotide-disulphide oxidoreductase dimerisation region [Silicibacter sp. TM1040] 101 ZP_010 . . . 86140117 soluble pyridine nucleotide transhydrogenase [Roseobacter sp. MED193] 102 BAB53939 14027670 soluble pyridine nucleotide transhydrogenase [Mesorhizobium loti MAFF303099] 103 YP_630631 108762684 NAD(P) transhydrogenase (B-specific) [Myxococcus xanthus DK 1622] 104 YP_467034 86160249 FAD-dependent pyridine nucleotide-disulphide oxidoreductase [Anaeromyxobacter dehalogenans 2CP-C] 105 YP_007724 46446359 soluble pyridine nucleotide transhydrogenase [Candidatus Protochlamydia amoebophila UWE25] 106 YP_433307 83644872 Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzyme [Hahella chejuensis KCTC 2396] 107 YP_527988 90022161 soluble pyridine nucleotide transhydrogenase [Saccharophagus degradans 2-40] 108 ZP_005 . . . 67931474 FAD-dependent pyridine nucleotide-disulphide oxidoreductase:Pyridine nucleotide-disulphide oxidoreductase dimerisation region [Solibacter usitatus Ellin6076] 109 YP_575887 92116158 Dihydrolipoamide dehydrogenase [Nitrobacter hamburgensis X14] 110 NP_539063 17986429 dihydrolipoamide dehydrogenase [Brucella melitensis 16M] 111 NP_698895 23502768 dihydrolipoamide dehydrogenase [Brucella suis 1330] 112 YP_415258 82700684 Pyridine nucleotide-disulphide oxidoreductase, class-II:NAD binding site:Adrenodoxin reductase:Mercuric reductase:Pyridine nu . . . [Brucella melitensis biovar Abortus 2308] 113 ZP_011 . . . 88945707 Dihydrolipoamide dehydrogenase [Desulfotomaculum reducens MI-1] 114 YP_675936 110635728 dihydrolipoamide dehydrogenase [Mesorhizobium sp. BNC1] 115 YP_483896 86747400 Dihydrolipoamide dehydrogenase [Rhodopseudomonas palustris HaA2] 116 ZP_006 . . . 71836831 Dihydrolipoamide dehydrogenase [Pelobacter propionicus DSM 2379] 117 BAB50985 14024381 ferric leghemoglobin reductase-2 precursor, dihydrolipoamide dehydrogenase [Mesorhizobium loti MAFF303099] 118 ZP_011 . . . 88946629 Dihydrolipoamide dehydrogenase [Desulfotomaculum reducens MI-1] 119 YP_032854 49474812 dihydrolipoamide dehydrogenase [Bartonella quintana str. Toulouse] 120 YP_317044 75674623 dihydrolipoamide dehydrogenase [Nitrobacter winogradskyi Nb- 255] 121 ZP_012 . . . 90421039 2-oxoglutarate dehydrogenase, E3 component, lipoamide dehydrogenase [Aurantimonas sp. SI85-9A1] 122 NP_533297 17936507 dihydrolipoamide dehydrogenase [Agrobacterium tumefaciens str. C58] 123 AAR21288 38489206 2-oxoglutarate dehydrogenase E3 component [Bartonella henselae] 124 YP_034342 49476301 dihydrolipoamide dehydrogenase [Bartonella henselae str. Houston-1] 125 ZP_008 . . . 78694497 Dihydrolipoamide dehydrogenase [Bradyrhizobium sp. BTAi1] 126 ZP_010 . . . 85713709 dihydrolipoamide dehydrogenase [Nitrobacter sp. Nb-311A] 127 YP_471355 86359463 dihydrolipoamide dehydrogenase protein [Rhizobium etli CFN 42] 128 ZP_008 . . . 77739670 Dihydrolipoamide dehydrogenase [Rhodopseudomonas palustris BisA53] 129 NP_387154 15966801 dihydrolipoamide dehydrogenase [Sinorhizobium meliloti 1021] 130 YP_567689 91975030 Dihydrolipoamide dehydrogenase [Rhodopseudomonas palustris BisB5] 131 ZP_009 . . . 83750811 COG1249: Pyruvate/2-oxoglutarate dehydrogenase complex, dihydrolipoamide dehydrogenase (E3) component, and related enzymes [Bartonella bacilliformis KC583] 132 NP_767089 27375560 dihydrolipoamide dehydrogenase [Bradyrhizobium japonicum USDA 110] 133 ZP_014 . . . 113873546 dihydrolipoamide dehydrogenase [Sinorhizobium medicae WSM419] 134 ZP_011 . . . 88801370 Pyruvate/2-oxoglutarate dehydrogenase complex [Polaribacter irgensii 23-P] 135 AAN03817 22652790 dihydrolipoamide dehydrogenase [Methylobacterium extorquens] 136 NP_945538 39933262 dihydrolipoamide dehydrogenase [Rhodopseudomonas palustris CGA009] 137 ZP_010 . . . 86142639 Dihydrolipoyl dehydrogenase [Flavobacterium sp. MED217] 138 YP_005669 46200002 dihydrolipoamide dehydrogenase [Thermus thermophilus HB27] 139 ZP_012 . . . 90589077 Dihydrolipoamide dehydrogenase [Flavobacterium johnsoniae UW101] 140 YP_143553 55980256 2-oxoglutarate dehydrogenase E3 component (dihydrolipoamide dehydrogenase) [Thermus thermophilus HB8] 141 YP_679939 110639729 dihydrolipoyl dehydrogenanse [Cytophaga hutchinsonii ATCC 33406] 142 YP_530077 90421707 Dihydrolipoamide dehydrogenase [Rhodopseudomonas palustris BisB18] 143 YP_681916 110678909 dihydrolipoamide dehydrogenase [Roseobacter denitrificans OCh 114] 144 ZP_010 . . . 86135035 dihydrolipoamide dehydrogenase [Tenacibaculum sp. MED152] 145 ZP_011 . . . 89068141 2-oxoglutarate dehydrogenase, E3 component, dihydrolipoamide dehydrogenase [Oceanicola granulosus HTCC2516] 146 ZP_007 . . . 75761004 Dihydrolipoamide dehydrogenase [Bacillus thuringiensis serovar israelensis ATCC 35646] 147 ZP_010 . . . 84684243 2-oxoglutarate dehydrogenase, E3 component, dihydrolipoamidedehydrogenase [Rhodobacterales bacterium HTCC2654] 148 ZP_009 . . . 84500199 2-oxoglutarate dehydrogenase, E3 component, dihydrolipoamidedehydrogenase [Oceanicola batsensis HTCC2597] 149 ZP_006 . . . 71900956 Dihydrolipoamide dehydrogenase [Xylella fastidiosa Ann-1] 150 XP_829730 74026328 dihydrolipoyl dehydrogenase [Trypanosoma brucei TREU927] 151 ZP_010 . . . 86132778 dihydrolipoamide dehydrogenase [Cellulophaga sp. MED134] 152 P71317 11135239 Soluble pyridine nucleotide transhydrogenase (STH) (NAD(P)(+) transhydrogenase [B-specific]) 153 ZP_012 . . . 89891405 dihydrolipoyl dehydrogenase (pyruvate 2-oxoglutarate dehydrogenase) [Flavobacteria bacterium BBFL7] 154 ZP_009 . . . 83950445 2-oxoglutarate dehydrogenase, E3 component, dihydrolipoamidedehydrogenase [Roseovarius nubinhibens ISM] 155 AAA21748 472330 dihydrolipoamide dehydrogenase 156 Q04933 416906 Dihydrolipoyl dehydrogenase (Dihydrolipoamide dehydrogenase) 157 ZP_011 . . . 88805521 Pyruvate/2-oxoglutarate dehydrogenase complex [Robiginitalea biformata HTCC2501] 158 YP_351398 77461891 dihydrolipoamide dehydrogenase [Pseudomonas fluorescens PfO-1] 159 ZP_006 . . . 71900678 Dihydrolipoamide dehydrogenase [Xylella fastidiosa Ann-1] 160 ZP_011 . . . 88936095 Dihydrolipoamide dehydrogenase [Geobacter uraniumreducens Rf4] 161 YP_430607 83590598 Dihydrolipoamide dehydrogenase [Moorella thermoacetica ATCC 39073] 162 YP_165603 56695257 2-oxoglutarate dehydrogenase, E3 component, dihydrolipoamide dehydrogenase [Silicibacter pomeroyi DSS-3] 163 ZP_011 . . . 89359154 Dihydrolipoamide dehydrogenase [Xanthobacter autotrophicus Py2] 164 YP_007150 46445785 dihydrolipoamide dehydrogenase [Candidatus Protochlamydia amoebophila UWE25] 165 YP_632394 108758911 alpha keto acid dehydrogenase complex, E3 component, lipoamide dehydrogenase [Myxococcus xanthus DK 1622] 166 ZP_010 . . . 85704799 2-oxoglutarate dehydrogenase, E3 component, dihydrolipoamidedehydrogenase [Roseovarius sp. 217] 167 NP_778978 28198664 dihydrolipoamide dehydrogenase [Xylella fastidiosa Temecula1] 168 ZP_011 . . . 88949385 Dihydrolipoamide dehydrogenase [Halorhodospira halophila SL1] 169 XP_749345 70988990 dihydrolipoamide dehydrogenase [Aspergillus fumigatus Af293] 170 YP_359567 78044229 alpha keto acid dehydrogenase complex, E3 component, lipoamide dehydrogenase [Carboxydothermus hydrogenoformans Z-2901] 171 XP_959535 85092766 hypothetical protein [Neurospora crassa OR74A] 172 ZP_009 . . . 83856665 Dihydrolipoyl dehydrogenase [Croceibacter atlanticus HTCC2559] 173 ZP_006 . . . 69259929 Dihydrolipoamide dehydrogenase [Magnetococcus sp. MC-1] 174 NP_298837 15838149 dihydrolipoamide dehydrogenase [Xylella fastidiosa 9a5c] 175 YP_508779 89053328 Dihydrolipoamide dehydrogenase [Jannaschia sp. CCS1] 176 CAA72131 1854569 dihydrolipoamide dehydrogenase [Trypanosoma cruzi] 177 ZP_011 . . . 88713490 Dihydrolipoyl dehydrogenase [Flavobacteriales bacterium HTCC2170] 178 P90597 6166121 Dihydrolipoyl dehydrogenase (Dihydrolipoamide dehydrogenase) 179 ZP_009 . . . 83944432 2-oxoglutarate dehydrogenase, E3 component, dihydrolipoamidedehydrogenase [Sulfitobacter sp. EE-36] 180 ZP_009 . . . 83953473 2-oxoglutarate dehydrogenase, E3 component, dihydrolipoamidedehydrogenase [Sulfitobacter sp. NAS-14.1] 181 ZP_010 . . . 86141935 dihydrolipoamide dehydrogenase, E3 component of 2- oxoglutarate and pyruvate dehydrogenase complexes [Flavobacterium sp. MED217] 182 XP_818792 71663602 dihydrolipoyl dehydrogenase [Trypanosoma cruzi strain CL Brener] 183 XP_812294 71422952 dihydrolipoyl dehydrogenase [Trypanosoma cruzi strain CL Brener] 184 CAA72132 1854571 dihydrolipoamide dehydrogenase [Trypanosoma cruzi] 185 NP_623271 20808100 Dihydrolipoamide dehydrogenase/glutathione oxidoreductase and related enzyme [Thermoanaerobacter tengcongensis MB4] 186 YP_265659 71082940 Dihydrolipoyl dehydrogenase [Candidatus Pelagibacter ubique HTCC1062] 187 ZP_012 . . . 91762636 Dihydrolipoyl dehydrogenase [Candidatus Pelagibacter ubique HTCC1002] 188 CAA11554 4210334 2-oxoglutarate dehydrogenase, E3 subunit [Arabidopsis thaliana] 189 NP_566570 30684428 LPD2 (LIPOAMIDE DEHYDROGENASE 2); FAD binding/ dihydrolipoyl dehydrogenase/disulfide oxidoreductase/ oxidoreductase [Arabidopsis thaliana] 190 ZP_010 . . . 84515869 2-oxoglutarate dehydrogenase, E3 component, dihydrolipoamidedehydrogenase [Loktanella vestfoldensis SKA53] 191 YP_355777 77917962 2-oxoglutarate dehydrogenase complex, E3 component, lipoamide dehydrogenase [Pelobacter carbinolicus DSM 2380] 192 AAS47708 44804791 dihydrolipoamide dehydrogenase [Leishmania major] 193 AAC26053 3309269 ferric leghemoglobin reductase-2 precursor [Glycine max] 194 XP_712334 68487571 putative mitochondrial matrix dihydrolipoamide dehydrogenase Lpd1p [Candida albicans SC5314] 195 ZP_008 . . . 78702525 Dihydrolipoamide dehydrogenase [Alkalilimnicola ehrlichei MLHE-1] 196 YP_002403 45658317 dihydrolipoamide dehydrogenase [Leptospira interrogans serovar Copenhageni str. Fiocruz L1-130] 197 ZP_008 . . . 82736566 Dihydrolipoamide dehydrogenase [Pseudomonas putida F1] 198 CAD60736 27803033 unnamed protein product [Podospora anserina] 199 NP_747467 26992042 dihydrolipoamide dehydrogenase [Pseudomonas putida KT2440] 200 ZP_013 . . . 106891371 Dihydrolipoamide dehydrogenase [Roseiflexus sp. RS-1] 201 EAQ92192 88184724 conserved hypothetical protein [Chaetomium globosum CBS 148.51]

TABLE 32 Examples of transhydrogenase subunit alpha polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 NP_416120 16129561 NAD(P) transhydrogenase subunit alpha [Escherichia coli K12]. 2 ZP_007 . . . 75512964 COG3288: NAD/NADP transhydrogenase alpha subunit [Escherichia coli 53638] 3 ZP_009 . . . 83587938 COG3288: NAD/NADP transhydrogenase alpha subunit [Escherichia coli 101-1] 4 YP_669455 110641725 NAD(P) transhydrogenase subunit alpha [Escherichia coli 536] 5 YP_540800 91210814 NAD(P) transhydrogenase subunit alpha [Escherichia coli UTI89] 6 ZP_007 . . . 75236686 COG3288: NAD/NADP transhydrogenase alpha subunit [Escherichia coli F11] 7 NP_753890 26247850 NAD(P) transhydrogenase subunit alpha [Escherichia coli CFT073] 8 YP_310491 74312072 pyridine nucleotide transhydrogenase, alpha subunit [Shigella sonnei Ss046] 9 ZP_007 . . . 75234916 COG3288: NAD/NADP transhydrogenase alpha subunit [Escherichia coli E110019] 10 YP_403177 82776828 pyridine nucleotide transhydrogenase, alpha subunit [Shigella dysenteriae Sd197] 11 ZP_007 . . . 75190643 COG3288: NAD/NADP transhydrogenase alpha subunit [Escherichia coli E24377A] 12 ZP_009 . . . 83570993 COG3288: NAD/NADP transhydrogenase alpha subunit [Shigella dysenteriae 1012] 13 ZP_007 . . . 75257022 COG3288: NAD/NADP transhydrogenase alpha subunit [Escherichia coli E22] 14 YP_407977 82544030 pyridine nucleotide transhydrogenase, alpha subunit [Shigella boydii Sb227] 15 ZP_006 . . . 75178502 COG3288: NAD/NADP transhydrogenase alpha subunit [Shigella boydii BS512] 16 YP_689096 110805576 pyridine nucleotide transhydrogenase, alpha subunit [Shigella flexneri 5 str. 8401] 17 NP_310336 15831563 NAD(P) transhydrogenase subunit alpha [Escherichia coli O157:H7 str. Sakai] 18 YP_216484 62180067 NAD(P) transhydrogenase subunit alpha [Salmonella enterica subsp. enterica serovar Choleraesuis str. SC-B67] 19 NP_460439 16764824 NAD(P) transhydrogenase subunit alpha [Salmonella typhimurium LT2] 20 NP_805194 29141852 NAD(P) transhydrogenase subunit alpha [Salmonella enterica subsp. enterica serovar Typhi Ty2] 21 ABF71058 102621428 NADP transhydrogenase alpha subunit [Enterobacter cloacae] 22 ZP_008 . . . 77963333 COG3288: NAD/NADP transhydrogenase alpha subunit [Yersinia mollaretii ATCC 43969] 23 ZP_008 . . . 77958020 COG3288: NAD/NADP transhydrogenase alpha subunit [Yersinia bercovieri ATCC 43970] 24 ZP_008 . . . 77975377 COG3288: NAD/NADP transhydrogenase alpha subunit [Yersinia frederiksenii ATCC 33641] 25 YP_647693 108811926 NAD(P) transhydrogenase alpha subunit [Yersinia pestis Nepal516] 26 YP_070740 51596549 NAD(P) transhydrogenase subunit alpha [Yersinia pseudotuberculosis IP 32953] 27 ZP_007 . . . 77629299 COG3288: NAD/NADP transhydrogenase alpha subunit [Yersinia pseudotuberculosis IP 31758] 28 YP_050297 50121130 NAD(P) transhydrogenase subunit alpha [Erwinia carotovora subsp. atroseptica SCRI1043] 29 ZP_008 . . . 77979498 COG3288: NAD/NADP transhydrogenase alpha subunit [Yersinia intermedia ATCC 29909] 30 NP_929427 37526083 NAD(P) transhydrogenase subunit alpha [Photorhabdus luminescens subsp. laumondii TTO1] 31 YP_455168 85059466 pyridine nucleotide transhydrogenase alpha subunit [Sodalis glossinidius str. ‘morsitans’] 32 YP_206544 59713769 NAD(P) transhydrogenase subunit alpha [Vibrio fischeri ES114] 33 ZP_007 . . . 75831475 COG3288: NAD/NADP transhydrogenase alpha subunit [Vibrio cholerae MO10] 34 ZP_010 . . . 86147993 NAD(P) transhydrogenase subunit alpha [Vibrio sp. MED222] 35 ZP_009 . . . 84393928 NAD(P) transhydrogenase subunit alpha [Vibrio splendidus 12B01] 36 ZP_007 . . . 75857576 COG3288: NAD/NADP transhydrogenase alpha subunit [Vibrio sp. Ex25] 37 ZP_012 . . . 91226112 NAD(P) transhydrogenase subunit alpha [Vibrio alginolyticus 12G01] 38 NP_936868 37676472 NAD(P) transhydrogenase subunit alpha [Vibrio vulnificus YJ016] 39 NP_800432 28900777 NAD(P) transhydrogenase subunit alpha [Vibrio parahaemolyticus RIMD 2210633] 40 NP_762287 27366760 NAD(P) transhydrogenase subunit alpha [Vibrio vulnificus CMCP6] 41 ZP_012 . . . 90577759 NAD(P) transhydrogenase subunit alpha [Vibrio angustum S14] 42 ZP_011 . . . 89073747 NAD(P) transhydrogenase subunit alpha [Photobacterium sp. SKA34] 43 NP_245690 15602618 NAD(P) transhydrogenase subunit alpha [Pasteurella multocida subsp. multocida str. Pm70] 44 YP_088416 52425279 NAD(P) transhydrogenase subunit alpha [Mannheimia succiniciproducens MBEL55E] 45 YP_718440 113460378 NAD(P)(+) transhydrogenase, alpha subunit (pyridine nucleotide transhydrogenase) [Haemophilus somnus 129PT] 46 ZP_001 . . . 32030098 COG3288: NAD/NADP transhydrogenase alpha subunit [Haemophilus somnus 2336] 47 ZP_005 . . . 68545628 NAD(P) transhydrogenase, alpha subunit [Shewanella amazonensis SB2B] 48 ZP_012 . . . 90412639 NAD(P) transhydrogenase subunit alpha [Photobacterium profundum 3TCK] 49 ZP_007 . . . 75429914 NAD/NADP transhydrogenase alpha subunit [Actinobacillus succinogenes 130Z] 50 YP_249235 68250123 NAD(P) transhydrogenase subunit alpha [Haemophilus influenzae 86-028NP] 51 YP_132574 54302581 NAD(P) transhydrogenase subunit alpha [Photobacterium profundum SS9] 52 NP_439513 16273272 NAD(P) transhydrogenase subunit alpha [Haemophilus influenzae Rd KW20] 53 ZP_009 . . . 82741968 NAD(P) transhydrogenase, alpha subunit [Shewanella sp. W3- 18-1] 54 ZP_014 . . . 113949477 NAD(P) transhydrogenase, alpha subunit [Shewanella baltica OS195] 55 YP_561942 91792291 NAD(P) transhydrogenase, alpha subunit [Shewanella denitrificans OS217] 56 NP_719279 24375236 NAD(P) transhydrogenase subunit alpha [Shewanella oneidensis MR-1] 57 NP_874046 33152693 NAD(P) transhydrogenase subunit alpha [Haemophilus ducreyi 35000HP] 58 YP_735209 113971416 NAD(P) transhydrogenase, alpha subunit [Shewanella sp. MR- 4] 59 ZP_008 . . . 78686308 NAD(P) transhydrogenase, alpha subunit [Shewanella sp. ANA- 3] 60 YP_736947 114046397 NAD(P) transhydrogenase, alpha subunit [Shewanella sp. MR- 7] 61 ZP_006 . . . 69950756 NAD(P) transhydrogenase, alpha subunit [Shewanella frigidimarina NCIMB 400] 62 ZP_007 . . . 75818840 COG3288: NAD/NADP transhydrogenase alpha subunit [Vibrio cholerae V51] 63 ZP_013 . . . 106883669 NAD(P) transhydrogenase, alpha subunit [Psychromonas ingrahamii 37] 64 NP_283944 15794122 NAD(P) transhydrogenase subunit alpha [Neisseria meningitidis Z2491] 65 YP_434236 83645801 NAD(P) transhydrogenase, alpha subunit [Hahella chejuensis KCTC 2396] 66 NP_274017 15676872 NAD(P) transhydrogenase subunit alpha [Neisseria meningitidis MC58] 67 YP_208522 59801810 NAD(P) transhydrogenase subunit alpha [Neisseria gonorrhoeae FA 1090] 68 ZP_012 . . . 90408904 PntA [Psychromonas sp. CNPT3] 69 YP_268514 71281713 NAD(P) transhydrogenase subunit alpha [Colwellia psychrerythraea 34H] 70 ZP_001 . . . 32035329 COG3288: NAD/NADP transhydrogenase alpha subunit [Actinobacillus pleuropneumoniae serovar 1 str. 4074] 71 ZP_012 . . . 90202099 NAD(P) transhydrogenase, alpha subunit [Mycobacterium vanbaalenii PYR-1] 72 YP_342322 77163797 NAD(P) transhydrogenase subunit alpha [Nitrosococcus oceani ATCC 19707] 73 ZP_009 . . . 84495285 NAD(P) transhydrogenase subunit alpha [Janibacter sp. HTCC2649] 74 AAY87755 68165590 NAD(P) transhydrogenase alpha subunit [Vibrio cholerae] 75 ZP_011 . . . 89340414 NAD(P) transhydrogenase, alpha subunit [Mycobacterium flavescens PYR-GCK] 76 AAY87751 68165582 NAD(P) transhydrogenase alpha subunit [Vibrio cholerae] 77 AAY87754 68165588 NAD(P) transhydrogenase alpha subunit [Vibrio cholerae] 78 AAY87762 68165604 NAD(P) transhydrogenase alpha subunit [Vibrio cholerae] 79 YP_422187 83311923 NAD/NADP transhydrogenase alpha subunit [Magnetospirillum magneticum AMB-1] 80 YP_637279 108797082 NAD(P) transhydrogenase, alpha subunit [Mycobacterium sp. MCS] 81 YP_115165 53803088 NAD(P) transhydrogenase subunit alpha [Methylococcus capsulatus str. Bath] 82 NP_821578 29826944 NAD(P) transhydrogenase subunit alpha [Streptomyces avermitilis MA-4680] 83 ZP_006 . . . 69261416 NAD(P) transhydrogenase, alpha subunit [Magnetococcus sp. MC-1] 84 NP_440856 16330128 NAD(P) transhydrogenase subunit alpha [Synechocystis sp. PCC 6803] 85 ZP_002 . . . 46202292 COG3288: NAD/NADP transhydrogenase alpha subunit [Magnetospirillum magnetotacticum MS-1] 86 YP_298860 73538493 NAD(P) transhydrogenase subunit alpha [Ralstonia eutropha JMP134] 87 ZP_004 . . . 66966825 NAD(P) transhydrogenase, alpha subunit [Arthrobacter sp. FB24] 88 ZP_006 . . . 71366730 NAD(P) transhydrogenase, alpha subunit [Nocardioides sp. JS614] 89 YP_524985 89902514 NAD(P) transhydrogenase, alpha subunit [Rhodoferax ferrireducens T118] 90 YP_055339 50842112 NAD(P) transhydrogenase subunit alpha [Propionibacterium acnes KPA171202] 91 NP_631664 21225885 NAD(P) transhydrogenase subunit alpha [Streptomyces coelicolor A3(2)] 92 ZP_006 . . . 74018096 NAD(P) transhydrogenase, alpha subunit [Burkholderia ambifaria AMMD] 93 CAJ96500 113530153 NAD(P) transhydrogenase subunit alpha [Ralstonia eutropha H16] 94 YP_366916 78060341 NAD(P) transhydrogenase subunit alpha [Burkholderia sp. 383] 95 YP_574538 92114610 NAD(P) transhydrogenase, alpha subunit [Chromohalobacter salexigens DSM 3043] 96 ZP_004 . . . 67542770 NAD(P) transhydrogenase, alpha subunit [Burkholderia vietnamiensis G4] 97 YP_702683 111019711 NAD(P) transhydrogenase alpha subunit [Rhodococcus sp. RHA1] 98 ZP_007 . . . 75821435 COG3288: NAD/NADP transhydrogenase alpha subunit [Vibrio cholerae RC385] 99 ZP_011 . . . 88798022 NAD(P) transhydrogenase subunit alpha [Reinekea sp. MED297] 100 ZP_009 . . . 83371049 NAD(P) transhydrogenase, alpha subunit [Rhodobacter sphaeroides ATCC 17025] 101 ZP_006 . . . 69933435 NAD(P) transhydrogenase, alpha subunit [Paracoccus denitrificans PD1222] 102 YP_353313 77463809 NAD(P) transhydrogenase subunit alpha [Rhodobacter sphaeroides 2.4.1] 103 ZP_009 . . . 83373326 NAD(P) transhydrogenase, alpha subunit [Rhodobacter sphaeroides ATCC 17029] 104 ZP_010 . . . 85704984 NAD(P) transhydrogenase subunit alpha [Roseovarius sp. 217] 105 YP_612703 99080549 NAD(P) transhydrogenase, alpha subunit [Silicibacter sp. TM1040] 106 YP_683504 110680497 NAD(P) transhydrogenase, alpha subunit [Roseobacter denitrificans OCh 114] 107 AAQ87369 36958944 NAD(P) transhydrogenase subunit alpha [Rhizobium sp. NGR234] 108 ZP_010 . . . 86137634 NAD(P) transhydrogenase subunit alpha [Roseobacter sp. MED193] 109 XP_799127 72030333 PREDICTED: similar to nicotinamide nucleotide transhydrogenase [Strongylocentrotus purpuratus] 110 ZP_010 . . . 84503572 NAD(P) transhydrogenase subunit alpha [Oceanicola batsensis HTCC2597] 111 ZP_010 . . . 84516763 NAD(P) transhydrogenase subunit alpha [Loktanella vestfoldensis SKA53] 112 NP_533164 17936374 NAD(P) transhydrogenase subunit alpha [Agrobacterium tumefaciens str. C58] 113 XP_562937 57968100 ENSANGP00000025507 [Anopheles gambiae str. PEST] 114 YP_511224 89055773 NAD(P) transhydrogenase, alpha subunit [Jannaschia sp. CCS1] 115 ZP_009 . . . 83942708 NAD(P) transhydrogenase subunit alpha [Sulfitobacter sp. EE- 36] 116 YP_168028 56697658 NAD(P) transhydrogenase subunit alpha [Silicibacter pomeroyi DSS-3] 117 EAT35199 108870974 nadp transhydrogenase [Aedes aegypti] 118 ZP_009 . . . 83953947 NAD(P) transhydrogenase subunit alpha [Sulfitobacter sp. NAS- 14.1] 119 ZP_009 . . . 83949483 NAD(P) transhydrogenase subunit alpha [Roseovarius nubinhibens ISM] 120 XP_367628 39972475 hypothetical protein MG07539.4 [Magnaporthe grisea 70-15] 121 NP_999921 47550793 nicotinamide nucleotide transhydrogenase [Danio rerio] 122 XP_961047 85100871 hypothetical protein (mitochondrial nicotinamide nucleotide transhydrogenase-related protein [imported] - Neurospora crassa OR74A) 123 AAH81117 51703526 MGC83563 protein [Xenopus laevis] 124 CAF99322 47223713 unnamed protein product [Tetraodon nigroviridis] 125 CAB88572 16416047 related to mitochondrial nicotinamide nucleotide transhydrogenase [Neurospora crassa] 126 AAI10544 83405154 Nicotinamide nucleotide transhydrogenase [Homo sapiens] 127 AAI10545 83405537 Nicotinamide nucleotide transhydrogenase [Homo sapiens] 128 NP_509028 17550456 Nicotinamide Nucleotide Transhydrogenase family member (nnt-1) [Caenorhabditis elegans] 129 2211247B 1589396 nicotinamide nucleotide transhydrogenase 130 ZP_010 . . . 84686399 NAD(P) transhydrogenase subunit alpha [Rhodobacterales bacterium HTCC2654] 131 NP_776368 27806831 nicotinamide nucleotide transhydrogenase [Bos taurus] 132 AAA21440 163397 nicotinamide nucleotide transhydrogenase 133 P11024 128400 NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) 134 XP_536481 73953769 PREDICTED: similar to NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) isoform 1 [Canis familiaris] 135 XP_970382 91083631 PREDICTED: similar to nicotinamide nucleotide transhydrogenase [Tribolium castaneum] 136 NP_001 . . . 61557127 nicotinamide nucleotide transhydrogenase [Rattus norvegicus] 137 NP_892022 33695086 nicotinamide nucleotide transhydrogenase [Homo sapiens] 138 XP_867928 73953777 PREDICTED: similar to NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) isoform 5 [Canis familiaris] 139 NP_036475 33695084 nicotinamide nucleotide transhydrogenase [Homo sapiens] 140 XP_424784 50761621 PREDICTED: similar to nicotinamide nucleotide transhydrogenase [Gallus gallus] 141 XP_646840 66826971 NAD(P)+ transhydrogenase (AB-specific) [Dictyostelium discoideum AX4] 142 BAC40113 26352966 unnamed protein product [Mus musculus] 143 EAS31570 90301939 conserved hypothetical protein [Coccidioides immitis RS] 144 XP_867922 73953775 PREDICTED: similar to NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) isoform 4 [Canis familiaris] 145 CAF99856 47230663 unnamed protein product [Tetraodon nigroviridis] 146 Q61941 51338804 NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) 147 2211247A 1589395 nicotinamide nucleotide transhydrogenase 148 NP_032736 31543330 nicotinamide nucleotide transhydrogenase [Mus musculus] 149 BAE35294 74198248 unnamed protein product [Mus musculus] 150 XP_748064 70985116 nicotinamide nucleotide transhydrogenase [Aspergillus fumigatus Af293] 151 XP_389182 46130902 hypothetical protein FG09006.1 [Gibberella zeae PH-1] 152 CAE68875 39598183 Hypothetical protein CBG14838 [Caenorhabditis briggsae] 153 AAK18179 13310152 nicotinamide nucleotide transhydrogenase [Acetabularia acetabulum] 154 AAG02246 11597233 NAD(P)H transhydrogenase [Acetabularia acetabulum] 155 AAC41577 6478876 pyridine nucleotidetranshydrogenase [Entamoeba histolytica] 156 XP_627136 66359916 pyridine nucleotide/NAD(P) transhydrogenase alpha plus beta subunits, duplicated gene, [Cryptosporidium parvum Iowa II] 157 XP_666495 67602649 transhydrogenase 7B2 [Cryptosporidium hominis TU502] 158 BAE40577 74219995 unnamed protein product [Mus musculus] 159 CAH90079 55726630 hypothetical protein [Pongo pygmaeus] 160 AAA80188 571367 nicotinamide nucleotide transhydrogenase 161 EAQ86424 88178956 conserved hypothetical protein [Chaetomium globosum CBS 148.51] 162 AAA29077 305062 transhydrogenase 163 AAA29081 158904 transhydrogenase 164 YP_724141 113478080 NAD(P)(+) transhydrogenase (AB-specific) [Trichodesmium erythraeum IMS101] 165 NP_487450 17230902 nicotinamide nucleotide transhydrogenase, subunit alpha [Nostoc sp. PCC 7120] 166 YP_323932 75909636 Alanine dehydrogenase/PNT-like [Anabaena variabilis ATCC 29413] 167 ZP_005 . . . 67924536 NAD(P)(+) transhydrogenase (AB-specific) [Crocosphaera watsonii WH 8501] 168 ZP_013 . . . 110607624 Alanine dehydrogenase/PNT-like [Maricaulis maris MCS10] 169 ZP_012 . . . 91222948 NAD(P) transhydrogenase subunit alpha [Psychroflexus torquis ATCC 700755] 170 ZP_009 . . . 82737423 NAD/NADP transhydrogenase, NAD(H)-binding DI subunit [Pseudomonas putida F1] 171 YP_000046 45655960 proton-translocating transhydrogenase, subunit alpha part 1 [Leptospira interrogans serovar Copenhageni str. Fiocruz L1- 130] 172 YP_257267 70733627 NAD(P) transhydrogenase, alpha subunit part 1 [Pseudomonas fluorescens Pf-5] 173 ZP_001 . . . 23130320 COG3288: NAD/NADP transhydrogenase alpha subunit [Nostoc punctiforme PCC 73102] 174 ZP_013 . . . 110593183 NAD(P)(+) transhydrogenase (AB-specific) [Acidovorax sp. JS42] 175 ZP_009 . . . 83859916 nicotinamide nucleotide transhydrogenase, subunit [Oceanicaulis alexandrii HTCC2633] 176 YP_526430 90020603 Glycogen/starch/alpha-glucan phosphorylase [Saccharophagus degradans 2-40] 177 ZP_012 . . . 89362826 nicotinamide nucleotide transhydrogenase, subunit alpha1 [Xanthobacter autotrophicus Py2] 178 YP_277175 71737873 NAD(P) transhydrogenase, subunit alpha part 1 [Pseudomonas syringae pv. phaseolicola 1448A] 179 YP_238081 66048240 Alanine dehydrogenase/PNT, C-terminal:Alanine dehydrogenase/PNT, N-terminal [Pseudomonas syringae pv. syringae B728a] 180 YP_675347 110635139 alanine dehydrogenase/PNT-like [Mesorhizobium sp. BNC1] 181 YP_345847 77456342 NAD(P)(+) transhydrogenase (AB-specific) [Pseudomonas fluorescens PfO-1] 182 AAQ87237 36958769 NAD(P) transhydrogenase subunit alpha [Rhizobium sp. NGR234] 183 ZP_013 . . . 94499798 pyridine nucleotide transhydrogenase alpha subunit [Oceanobacter sp. RED65] 184 NP_881481 33593837 NAD(P) transhydrogenase, subunit alpha part 1 [Bordetella pertussis Tohama I] 185 P0C186 91207608 NAD(P) transhydrogenase subunit alpha part 1 (Pyridine nucleotide transhydrogenase subunit alpha 1) (Nicotinamide nucleotide transhydrogenase subunit alpha 1) (Proton- translocating transhydrogenase component 1) (dI) 186 1PTJB 37927621 Chain B, Crystal Structure Analysis Of The Di And Diii Complex Of Transhydrogenase With A Thio-Nicotinamide Nucleotide Analogue 187 YP_485056 86748560 NAD(P)(+) transhydrogenase (AB-specific) [Rhodopseudomonas palustris HaA2] 188 YP_620103 107021776 alanine dehydrogenase/PNT-like [Burkholderia cenocepacia AU 1054] 189 ZP_008 . . . 77952426 pyridine nucleotide transhydrogenase alpha subunit [Marinobacter aquaeolei VT8] 190 NP_840933 30248863 Alanine dehydrogenase and pyridine nucleotide transhydrogenase [Nitrosomonas europaea ATCC 19718] 191 2FRDB 90109601 Chain B, Structure Of Transhydrogenase (Di.S138a.Nadh)2(Diii.Nadph)1 Asymmetric Complex 192 NP_888491 33600931 NAD(P) transhydrogenase, subunit alpha part 1 [Bordetella bronchiseptica RB50] 193 XP_726447 82596885 NAD(P) transhydrogenase subunit beta [Plasmodium yoelii yoelii str. 17XNL] 194 YP_551329 91790377 NAD(P)(+) transhydrogenase (AB-specific) [Polaromonas sp. JS666] 195 ZP_006 . . . 71548985 NAD(P)(+) transhydrogenase (AB-specific) [Nitrosomonas eutropha C71] 196 YP_568553 91975894 NAD(P)(+) transhydrogenase (AB-specific) [Rhodopseudomonas palustris BisB5] 197 2FSVB 90109621 Chain B, Structure Of Transhydrogenase (Di.D135n.Nad+) 2(Diii.E155w.Nadp+)1 Asymmetric Complex 198 1NM5B 42543078 Chain B, R. Rubrum Transhydrogenase (Di.Q132n)2(Diii)1 Asymmetric Complex 199 1L7DB 27065343 Chain B, Crystal Structure Of R. Rubrum Transhydrogenase Domain I Without Bound Nad(H) 200 ZP_012 . . . 90418769 NAD(P) transhydrogenase, subunit alpha part 1 [Aurantimonas sp. SI85-9A1] 201 2FR8B 90109598 Chain B, Structure Of Transhydrogenase (Di.R127a.Nad+)2(Diii.Nadp+)1 Asymmetric Complex

TABLE 33 Examples of transhydrogenase subunit beta polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 NP_416119 16129560 pyridine nucleotide transhydrogenase [Escherichia coli K12]. 2 YP_669454 110641724 NAD(P) transhydrogenase subunit beta [Escherichia coli 536] 3 ZP_007 . . . 75234915 COG1282: NAD/NADP transhydrogenase beta subunit [Escherichia coli E110019] 4 ZP_007 . . . 75230949 COG1282: NAD/NADP transhydrogenase beta subunit [Escherichia coli B7A] 5 YP_407978 82544031 pyridine nucleotide transhydrogenase, beta subunit [Shigella boydii Sb227] 6 YP_150630 56413555 pyridine nucleotide transhydrogenase [Salmonella enterica subsp. enterica serovar Paratyphi A str. ATCC 9150] 7 YP_216485 62180068 pyridine nucleotide transhydrogenase [Salmonella enterica subsp. enterica serovar Choleraesuis str. SC-B67] 8 ABF71059 102621429 NADP transhydrogenase beta subunit [Enterobacter cloacae] 9 NP_929428 37526084 pyridine nucleotide transhydrogenase [Photorhabdus luminescens subsp. laumondii TTO1] 10 YP_050298 50121131 pyridine nucleotide transhydrogenase [Erwinia carotovora subsp. atroseptica SCRI1043] 11 ZP_008 . . . 77979497 COG1282: NAD/NADP transhydrogenase beta subunit [Yersinia intermedia ATCC 29909] 12 ZP_008 . . . 77975378 COG1282: NAD/NADP transhydrogenase beta subunit [Yersinia frederiksenii ATCC 33641] 13 ZP_008 . . . 77958019 COG1282: NAD/NADP transhydrogenase beta subunit [Yersinia bercovieri ATCC 43970] 14 ZP_008 . . . 77963332 COG1282: NAD/NADP transhydrogenase beta subunit [Yersinia mollaretii ATCC 43969] 15 YP_647692 108811925 NAD(P) transhydrogenase subunit beta [Yersinia pestis Nepal516] 16 YP_455167 85059465 pyridine nucleotide transhydrogenase beta subunit [Sodalis glossinidius str. ‘morsitans’] 17 NP_719280 24375237 pyridine nucleotide transhydrogenase [Shewanella oneidensis MR-1] 18 YP_736946 114046396 NAD(P) transhydrogenase, beta subunit [Shewanella sp. MR-7] 19 ZP_009 . . . 82741967 NAD(P) transhydrogenase, beta subunit [Shewanella sp. W3-18- 1] 20 NP_245689 15602617 pyridine nucleotide transhydrogenase [Pasteurella multocida subsp. multocida str. Pm70] 21 ZP_005 . . . 68541685 NAD(P) transhydrogenase, beta subunit [Shewanella baltica OS155] 22 ZP_012 . . . 90412640 putative NAD(P) transhydrogenase, beta subunit [Photobacterium profundum 3TCK] 23 ZP_001 . . . 53732668 COG1282: NAD/NADP transhydrogenase beta subunit [Haemophilus influenzae R2846] 24 YP_249234 68250122 pyridine nucleotide transhydrogenase [Haemophilus influenzae 86-028NP] 25 NP_439514 16273273 pyridine nucleotide transhydrogenase [Haemophilus influenzae Rd KW20] 26 ZP_005 . . . 68545629 NAD(P) transhydrogenase, beta subunit [Shewanella amazonensis SB2B] 27 YP_132575 54302582 pyridine nucleotide transhydrogenase [Photobacterium profundum SS9] 28 ZP_011 . . . 89073748 pyridine nucleotide transhydrogenase [Photobacterium sp. SKA34] 29 NP_800431 28900776 pyridine nucleotide transhydrogenase [Vibrio parahaemolyticus RIMD 2210633] 30 ZP_012 . . . 90577760 pyridine nucleotide transhydrogenase [Vibrio angustum S14] 31 ZP_007 . . . 75857577 COG1282: NAD/NADP transhydrogenase beta subunit [Vibrio sp. Ex25] 32 NP_936867 37676471 pyridine nucleotide transhydrogenase [Vibrio vulnificus YJ016] 33 ZP_007 . . . 75818461 COG1282: NAD/NADP transhydrogenase beta subunit [Vibrio cholerae V51] 34 ZP_007 . . . 75831476 COG1282: NAD/NADP transhydrogenase beta subunit [Vibrio cholerae MO10] 35 ZP_012 . . . 91226113 pyridine nucleotide transhydrogenase [Vibrio alginolyticus 12G01] 36 YP_206543 59713768 pyridine nucleotide transhydrogenase [Vibrio fischeri ES114] 37 YP_088415 52425278 pyridine nucleotide transhydrogenase [Mannheimia succiniciproducens MBEL55E] 38 ZP_001 . . . 32035330 COG1282: NAD/NADP transhydrogenase beta subunit [Actinobacillus pleuropneumoniae serovar 1 str. 4074] 39 ZP_006 . . . 69950757 NAD(P) transhydrogenase, beta subunit [Shewanella frigidimarina NCIMB 400] 40 ZP_010 . . . 86147992 pyridine nucleotide transhydrogenase [Vibrio sp. MED222] 41 ZP_009 . . . 84393927 NAD(P) transhydrogenase, beta subunit [Vibrio splendidus 12B01] 42 ZP_007 . . . 75429915 NAD/NADP transhydrogenase beta subunit [Actinobacillus succinogenes 130Z] 43 NP_274015 15676870 pyridine nucleotide transhydrogenase [Neisseria meningitidis MC58] 44 NP_283942 15794120 pyridine nucleotide transhydrogenase [Neisseria meningitidis Z2491] 45 YP_208524 59801812 pyridine nucleotide transhydrogenase [Neisseria gonorrhoeae FA 1090] 46 YP_718441 113460379 NAD(P)(+) transhydrogenase, beta subunit (pyridine nucleotide transhydrogenase) [Haemophilus somnus 129PT] 47 NP_874045 33152692 pyridine nucleotide transhydrogenase [Haemophilus ducreyi 35000HP] 48 YP_561941 91792290 NAD(P) transhydrogenase, beta subunit [Shewanella denitrificans OS217] 49 YP_434235 83645800 NAD/NADP transhydrogenase beta subunit [Hahella chejuensis KCTC 2396] 50 YP_268515 71280969 pyridine nucleotide transhydrogenase [Colwellia psychrerythraea 34H] 51 ZP_013 . . . 106883670 NAD(P) transhydrogenase, beta subunit [Psychromonas ingrahamii 37] 52 ZP_009 . . . 84495284 pyridine nucleotide transhydrogenase [Janibacter sp. HTCC2649] 53 YP_637278 108797081 NAD(P) transhydrogenase, beta subunit [Mycobacterium sp. MCS] 54 ZP_012 . . . 90408905 NAD(P) transhydrogenase subunit beta [Psychromonas sp. CNPT3] 55 ZP_011 . . . 89340413 NAD(P) transhydrogenase, beta subunit [Mycobacterium flavescens PYR-GCK] 56 ZP_012 . . . 90202100 NAD(P) transhydrogenase, beta subunit [Mycobacterium vanbaalenii PYR-1] 57 NP_631663 21225884 NAD(P) transhydrogenase beta subunit [Streptomyces coelicolor A3(2)] 58 NP_821577 29826943 pyridine nucleotide transhydrogenase, beta subunit [Streptomyces avermitilis MA-4680] 59 YP_702682 111019710 NAD(P) transhydrogenase beta subunit [Rhodococcus sp. RHA1] 60 YP_422186 83311922 NAD/NADP transhydrogenase beta subunit [Magnetospirillum magneticum AMB-1] 61 ZP_006 . . . 71366731 NAD(P) transhydrogenase, beta subunit [Nocardioides sp. JS614] 62 YP_115164 53803089 NAD(P) transhydrogenase, beta subunit [Methylococcus capsulatus str. Bath] 63 YP_524984 89902513 NAD(P) transhydrogenase, beta subunit [Rhodoferax ferrireducens T118] 64 NP_440860 16330132 pyridine nucleotide transhydrogenase beta subunit [Synechocystis sp. PCC 6803] 65 ZP_004 . . . 66966824 NAD(P) transhydrogenase, beta subunit [Arthrobacter sp. FB24] 66 ZP_011 . . . 88798021 pyridine nucleotide transhydrogenase [Reinekea sp. MED297] 67 YP_342321 77163796 NAD(P) transhydrogenase, beta subunit [Nitrosococcus oceani ATCC 19707] 68 ZP_006 . . . 69261415 NAD(P) transhydrogenase, beta subunit [Magnetococcus sp. MC-1] 69 ZP_004 . . . 67542769 NAD(P) transhydrogenase, beta subunit [Burkholderia vietnamiensis G4] 70 YP_366915 78060340 NAD(P) transhydrogenase, beta subunit [Burkholderia sp. 383] 71 ZP_006 . . . 74018097 NAD(P) transhydrogenase, beta subunit [Burkholderia ambifaria AMMD] 72 YP_298859 73538492 NAD(P) transhydrogenase, beta subunit [Ralstonia eutropha JMP134] 73 YP_055340 50842113 NAD(P) transhydrogenase subunit beta [Propionibacterium acnes KPA171202] 74 ZP_009 . . . 83949482 NAD(P)+ transhydrogenase, beta subunit [Roseovarius nubinhibens ISM] 75 CAJ96499 113530152 NAD(P) transhydrogenase subunit beta [Ralstonia eutropha H16] 76 ZP_006 . . . 69933434 NAD(P) transhydrogenase, beta subunit [Paracoccus denitrificans PD1222] 77 YP_168027 56697657 NAD(P)+ transhydrogenase, beta subunit [Silicibacter pomeroyi DSS-3] 78 NP_533165 17936375 NAD(P)+ transhydrogenase beta chain [Agrobacterium tumefaciens str. C58] 79 AAQ87370 36958945 NAD(P) transhydrogenase subunit beta [Rhizobium sp. NGR234] 80 ZP_009 . . . 83942707 NAD(P)+ transhydrogenase, beta subunit [Sulfitobacter sp. EE- 36] 81 ZP_009 . . . 83953946 NAD(P)+ transhydrogenase, beta subunit [Sulfitobacter sp. NAS-14.1] 82 ZP_007 . . . 75813705 COG1282: NAD/NADP transhydrogenase beta subunit [Vibrio cholerae V52] 83 ZP_010 . . . 84503573 NAD(P)+ transhydrogenase, beta subunit [Oceanicola batsensis HTCC2597] 84 ZP_010 . . . 84686400 NAD(P)+ transhydrogenase, beta subunit [Rhodobacterales bacterium HTCC2654] 85 YP_612702 99080548 NAD(P) transhydrogenase, beta subunit [Silicibacter sp. TM1040] 86 ZP_010 . . . 84516762 NAD(P)+ transhydrogenase, beta subunit [Loktanella vestfoldensis SKA53] 87 ZP_010 . . . 86137633 NAD(P)+ transhydrogenase, beta subunit [Roseobacter sp. MED193] 88 YP_511223 89055772 NAD(P) transhydrogenase, beta subunit [Jannaschia sp. CCS1] 89 ZP_010 . . . 85704983 NAD(P)+ transhydrogenase, beta subunit [Roseovarius sp. 217] 90 ZP_007 . . . 75822802 COG1282: NAD/NADP transhydrogenase beta subunit [Vibrio cholerae RC385] 91 YP_683503 110680496 NAD(P) transhydrogenase, beta subunit [Roseobacter denitrificans OCh 114] 92 NP_949516 39937240 nicotinamide nucleotide transhydrogenase, subunit beta [Rhodopseudomonas palustris CGA009] 93 YP_418478 83269187 NAD(P) transhydrogenase, beta subunit [Brucella melitensis biovar Abortus 2308] 94 YP_485058 86748562 NAD(P) transhydrogenase, beta subunit [Rhodopseudomonas palustris HaA2] 95 ZP_009 . . . 83859918 nicotinamide nucleotide transhydrogenase, subunit beta [Oceanicaulis alexandrii HTCC2633] 96 ZP_008 . . . 77740973 NAD(P) transhydrogenase, beta subunit [Rhodopseudomonas palustris BisA53] 97 ZP_012 . . . 89362731 NAD(P) transhydrogenase, beta subunit [Xanthobacter autotrophicus Py2] 98 ZP_008 . . . 77814001 NAD(P) transhydrogenase, beta subunit [Shewanella putrefaciens CN-32] 99 ZP_010 . . . 84704197 PntB, NAD(P) transhydrogenase, beta subunit [Parvularcula bermudensis HTCC2503] 100 YP_533811 90425441 NAD(P) transhydrogenase, beta subunit [Rhodopseudomonas palustris BisB18] 101 P0C188 91207612 NAD(P) transhydrogenase subunit beta (Pyridine nucleotide transhydrogenase subunit beta) (Nicotinamide nucleotide transhydrogenase subunit beta) (Proton-translocating transhydrogenase NADP(H)-binding component) (dIII) 102 YP_568555 91975896 NAD(P) transhydrogenase, beta subunit [Rhodopseudomonas palustris BisB5] 103 ZP_012 . . . 90418771 NAD(P) transhydrogenase, subunit beta [Aurantimonas sp. SI85-9A1] 104 YP_526432 90020605 NAD(P) transhydrogenase, beta subunit [Saccharophagus degradans 2-40] 105 ZP_008 . . . 78699497 NAD(P)+ transhydrogenase beta chain [Bradyrhizobium sp. BTAi1] 106 XP_424784 50761621 PREDICTED: similar to nicotinamide nucleotide transhydrogenase [Gallus gallus] 107 XP_799127 72030333 PREDICTED: similar to nicotinamide nucleotide transhydrogenase [Strongylocentrotus purpuratus] 108 ZP_010 . . . 85716091 NAD/NADP transhydrogenase beta subunit [Nitrobacter sp. Nb- 311A] 109 YP_605919 104779421 pyridine nucleotide transhydrogenase, beta subunit [Pseudomonas entomophila L48] 110 YP_457359 85373297 PntB, NAD(P) transhydrogenase, beta subunit [Erythrobacter litoralis HTCC2594] 111 XP_970382 91083631 PREDICTED: similar to nicotinamide nucleotide transhydrogenase [Tribolium castaneum] 112 BAB51677 14025075 nicotinamide nucleotide transhydrogenase, subunit beta [Mesorhizobium loti MAFF303099] 113 ZP_012 . . . 94413536 hypothetical protein PaerP_01004711 [Pseudomonas aeruginosa PA7] 114 ZP_011 . . . 88938736 NAD/NADP transhydrogenase beta subunit-like [Acidiphilium cryptum JF-5] 115 NP_795188 28872569 NAD(P) transhydrogenase, beta subunit [Pseudomonas syringae pv. tomato str. DC3000] 116 YP_257265 70733625 NAD(P) transhydrogenase, beta subunit [Pseudomonas fluorescens Pf-5] 117 NP_773764 27382235 NAD(P)+ transhydrogenase beta chain [Bradyrhizobium japonicum USDA 110] 118 YP_576517 92116788 NAD(P) transhydrogenase, beta subunit [Nitrobacter hamburgensis X14] 119 ZP_013 . . . 107099186 hypothetical protein PaerPA_01000196 [Pseudomonas aeruginosa PACS2] 120 YP_557045 91781839 Putative NAD(P) transhydrogenase, beta subunit [Burkholderia xenovorans LB400] 121 ZP_009 . . . 82737421 pyridine nucleotide transhydrogenase, beta subunit [Pseudomonas putida F1] 122 YP_345845 77456340 NAD(P) transhydrogenase, beta subunit [Pseudomonas fluorescens PfO-1] 123 NP_999921 47550793 nicotinamide nucleotide transhydrogenase [Danio rerio] 124 YP_411848 82702282 NAD(P) transhydrogenase, beta subunit [Nitrosospira multiformis ATCC 25196] 125 CAF99856 47230663 unnamed protein product [Tetraodon nigroviridis] 126 YP_675345 110635137 NAD(P) transhydrogenase, beta subunit [Mesorhizobium sp. BNC1] 127 EAT35199 108870974 nadp transhydrogenase [Aedes aegypti] 128 YP_277177 71736851 NAD(P) transhydrogenase, beta subunit [Pseudomonas syringae pv. phaseolicola 1448A] 129 YP_263852 71065125 NAD(P) transhydrogenase, beta subunit [Psychrobacter arcticus 273-4] 130 NP_840935 30248865 NAD(P) transhydrogenase beta subunit [Nitrosomonas europaea ATCC 19718] 131 XP_867922 73953775 PREDICTED: similar to NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) isoform 4 [Canis familiaris] 132 XP_867916 73953773 PREDICTED: similar to NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) isoform 3 [Canis familiaris] 133 XP_536481 73953769 PREDICTED: similar to NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) isoform 1 [Canis familiaris] 134 XP_867928 73953777 PREDICTED: similar to NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) isoform 5 [Canis familiaris] 135 NP_001 . . . 61557127 nicotinamide nucleotide transhydrogenase [Rattus norvegicus] 136 YP_238083 66048242 NAD(P) transhydrogenase, beta subunit [Pseudomonas syringae pv. syringae B728a] 137 BAE40577 74219995 unnamed protein product [Mus musculus] 138 BAC39226 26351179 unnamed protein product [Mus musculus] 139 ZP_013 . . . 94494989 NAD(P) transhydrogenase, beta subunit [Sphingomonas sp. SKA58] 140 NP_032736 31543330 nicotinamide nucleotide transhydrogenase [Mus musculus] 141 BAC40113 26352966 unnamed protein product [Mus musculus] 142 ZP_004 . . . 67155631 NAD(P) transhydrogenase, beta subunit [Azotobacter vinelandii AvOP] 143 YP_579812 93005375 NAD(P) transhydrogenase, beta subunit [Psychrobacter cryohalolentis K5] 144 YP_297031 73542511 NAD(P) transhydrogenase, beta subunit [Ralstonia eutropha JMP134] 145 YP_585107 94311897 NAD(P) transhydrogenase, beta subunit [Ralstonia metallidurans CH34] 146 YP_497119 87199862 NAD(P) transhydrogenase, beta subunit [Novosphingobium aromaticivorans DSM 12444] 147 AAQ87239 36958771 NAD(P) transhydrogenase subunit beta [Rhizobium sp. NGR234] 148 NP_888489 33600929 NAD(P) transhydrogenase subunit beta [Bordetella bronchiseptica RB50] 149 Q61941 51338804 NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) 150 NP_776368 27806831 nicotinamide nucleotide transhydrogenase [Bos taurus] 151 NP_884728 33597085 NAD(P) transhydrogenase subunit beta [Bordetella parapertussis 12822] 152 AAA21440 163397 nicotinamide nucleotide transhydrogenase 153 YP_441808 83721084 NAD(P) transhydrogenase, beta subunit [Burkholderia thailandensis E264] 154 AAI10544 83405154 Nicotinamide nucleotide transhydrogenase [Homo sapiens] 155 NP_899769 34495554 NAD(P) transhydrogenase, beta subunit [Chromobacterium violaceum ATCC 12472] 156 NP_422097 16127533 NAD(P) transhydrogenase, beta subunit [Caulobacter crescentus CB15] 157 XP_312859 58383882 ENSANGP00000016622 [Anopheles gambiae str. PEST] 158 ZP_009 . . . 84360819 COG1282: NAD/NADP transhydrogenase beta subunit [Burkholderia dolosa AUO158] 159 ZP_014 . . . 113935785 NAD(P) transhydrogenase, beta subunit [Caulobacter sp. K31] 160 ZP_006 . . . 71548983 NAD(P) transhydrogenase, beta subunit [Nitrosomonas eutropha C71] 161 AAH81117 51703526 MGC83563 protein [Xenopus laevis] 162 CAH90079 55726630 hypothetical protein [Pongo pygmaeus] 163 BAE35294 74198248 unnamed protein product [Mus musculus] 164 ZP_014 . . . 113872300 probable NAD(P) transhydrogenase subunit beta transmembrane protein [Sinorhizobium medicae WSM419] 165 AAI10545 83405537 Nicotinamide nucleotide transhydrogenase [Homo sapiens] 166 NP_036475 33695084 nicotinamide nucleotide transhydrogenase [Homo sapiens] 167 YP_727574 113869085 NAD(P) transhydrogenase subunit beta [Ralstonia eutropha H16] 168 2211247B 1589396 nicotinamide nucleotide transhydrogenase 169 2211247A 1589395 nicotinamide nucleotide transhydrogenase 170 AAN62246 24461677 putative pyridine nucleotide transhydrogenase, beta subunit [Pseudomonas aeruginosa] 171 P11024 128400 NAD(P) transhydrogenase, mitochondrial precursor (Pyridine nucleotide transhydrogenase) (Nicotinamide nucleotide transhydrogenase) 172 NP_892022 33695086 nicotinamide nucleotide transhydrogenase [Homo sapiens] 173 NP_386966 15966613 PROBABLE NAD(P) TRANSHYDROGENASE SUBUNIT BETA TRANSMEMBRANE PROTEIN [Sinorhizobium meliloti 1021] 174 YP_299910 73539543 NAD(P) transhydrogenase, beta subunit [Ralstonia eutropha JMP134] 175 ZP_012 . . . 91769178 NAD(P) transhydrogenase, beta subunit [Psychrobacter sp. PRwf-1] 176 YP_293241 72383887 NAD(P) transhydrogenase, beta subunit [Ralstonia eutropha JMP134] 177 YP_299480 73539113 NAD(P) transhydrogenase, beta subunit [Ralstonia eutropha JMP134] 178 YP_293567 72384213 NAD(P) transhydrogenase, beta subunit [Ralstonia eutropha JMP134] 179 YP_725774 113867285 NAD(P) transhydrogenase subunit beta [Ralstonia eutropha H16] 180 YP_620105 107021778 NAD(P) transhydrogenase, beta subunit [Burkholderia cenocepacia AU 1054] 181 YP_725367 113866878 NAD(P) transhydrogenase subunit beta [Ralstonia eutropha H16] 182 NP_520854 17547452 PROBABLE TRANSMEMBRANE NADP TRANSHYDROGENASE (BETA SUBUNIT) OXIDOREDUCTASE PROTEIN [Ralstonia solanacearum GMI1000] 183 ZP_009 . . . 83746755 PntB [Ralstonia solanacearum UW551] 184 ZP_013 . . . 100235717 hypothetical protein Bpse4_03000891 [Burkholderia pseudomallei 406e] 185 YP_334763 76809085 NAD(P) transhydrogenase subunit beta [Burkholderia pseudomallei 1710b] 186 ZP_013 . . . 100917505 hypothetical protein Bmal10_03001254 [Burkholderia mallei 10399] 187 ZP_004 . . . 67667725 NAD(P) transhydrogenase, beta subunit [Burkholderia cenocepacia HI2424] 188 ZP_012 . . . 91762289 NAD(p) transhydrogenase subunit beta [Candidatus Pelagibacter ubique HTCC1002] 189 YP_471093 86359201 NAD(P)(+) transhydrogenase (AB-specific), beta subunit protein [Rhizobium etli CFN 42] 190 ZP_010 . . . 85707884 PntB, NAD(P) transhydrogenase, beta subunit [Erythrobacter sp. NAP1] 191 YP_293266 72383912 NAD(P) transhydrogenase, beta subunit [Ralstonia eutropha JMP134] 192 ZP_009 . . . 84356795 COG1282: NAD/NADP transhydrogenase beta subunit [Burkholderia cenocepacia PC184] 193 YP_266000 71083281 NAD(p) transhydrogenase subunit beta [Candidatus Pelagibacter ubique HTCC1062] 194 CAE68875 39598183 Hypothetical protein CBG14838 [Caenorhabditis briggsae] 195 YP_368033 78065264 NAD(P) transhydrogenase, beta subunit [Burkholderia sp. 383] 196 YP_616428 103486867 NAD(P) transhydrogenase, beta subunit [Sphingopyxis alaskensis RB2256] 197 YP_246075 67458451 NAD(p) transhydrogenase subunit beta [Rickettsia felis URRWXCal2] 198 YP_537194 91204839 NAD(p) transhydrogenase subunit beta [Rickettsia bellii RML369-C] 199 XP_646840 66826971 NAD(P)+ transhydrogenase (AB-specific) [Dictyostelium discoideum AX4] 200 ZP_009 . . . 83747817 NAD(P) transhydrogenase subunit beta [Ralstonia solanacearum UW551] 201 YP_047599 50086089 pyridine nucleotide transhydrogenase, beta subunit [Acinetobacter sp. ADP1]

TABLE 34 Examples of pox2 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_505264.1 50555712 YlPOX2 [Yarrowia lipolytica]. 2 O74936 59799074 Acyl-coenzyme A oxidase 3 (Acyl-CoA oxidase 3) 3 XP_504475 50554133 YlPOX4 [Yarrowia lipolytica] 4 XP_502199 50549457 YlPOX5 [Yarrowia lipolytica] 5 O74934 59799072 Acyl-coenzyme A oxidase 1 (Acyl-CoA oxidase 1) 6 XP_504703 50554589 YlPOX1 [Yarrowia lipolytica] 7 XP_503632 50552444 hypothetical protein [Yarrowia lipolytica] 8 XP_760471 71020481 hypothetical protein UM04324.1 [Ustilago maydis 521] 9 XP_571768 58269224 Acyl-coenzyme A oxidase I [Cryptococcus neoformans var. neoformans JEC21] 10 XP_717995 68475844 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 11 P08790 17373073 Acyl-coenzyme A oxidase 5 (Acyl-CoA oxidase 5) (PXP-5) 12 Q6BRD5 59799027 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 13 OXCKX5 66061 acyl-CoA oxidase (EC 1.3.3.6) POX5, peroxisomal - yeast (Candida tropicalis ) 14 BAA83482 5763520 acyl-CoA oxidase [Candida tropicalis] 15 XP_457726 50417804 hypothetical protein DEHA0C01155g [Debaryomyces hansenii CBS767] 16 BAA83483 5763522 acyl-CoA oxidase [Candida tropicalis] 17 OXCKPM 66060 acyl-CoA oxidase (EC 1.3.3.6) PXP4, peroxisomal - yeast (Candida maltosa ) 18 AAA34362 170912 acyl-coenzyme A oxidase II precursor 19 Q9Y7B1 60391213 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 20 XP_721610 68468582 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 21 AAA34361 170910 PXP-2 protein 22 1306283A 225549 oxidase, fatty acyl 23 P06598 17373023 Acyl-coenzyme A oxidase 4 (Acyl-CoA oxidase 4) (PXP-4) (Peroxisomal fatty acyl-CoA oxidase) 24 XP_721613 68468588 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 25 XP_459232 50421365 hypothetical protein DEHA0D18667g [Debaryomyces hansenii CBS767] 26 Q00468 2498206 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) (AOX 2) 27 Q6CKK7 59799028 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 28 NP_011310 6321233 Fatty-acyl coenzyme A oxidase, involved in the fatty acid beta- oxidation pathway; localized to the peroxisomal matrix; Pox1p [Saccharomyces cerevisiae] 29 AAA34891 172217 acyl-coenzyme A oxidase 30 Q6FY63 59799032 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 31 Q756A9 60391210 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 32 O65202 62286589 Acyl-coenzyme A oxidase 1, peroxisomal (AOX 1) (Long-chain acyl-CoA oxidase) (AtCX1) 33 1W07B 58177067 Chain B, Arabidopsis Thaliana Acyl-Coa Oxidase 1 34 NP_567513 18414744 ACX1 (ACYL-COA OXIDASE 1) [Arabidopsis thaliana] 35 H71434 7488059 probable apetala2 domain TINY - Arabidopsis thaliana 36 XP_001 . . . 109118312 PREDICTED: similar to acyl-Coenzyme A oxidase isoform b [Macaca mulatta] 37 Q5RC19 62286600 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 38 Q15067 17380467 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) (Straight-chain acyl-CoA oxidase) (SCOX) 39 NP_009223 30089974 acyl-Coenzyme A oxidase isoform b [Homo sapiens] 40 AAB30019 7689914 peroxisomal acyl-coenzyme A oxidase [Homo sapiens] 41 CAH90691 55727877 hypothetical protein [Pongo pygmaeus] 42 AAL01888 15553480 acyl-CoA oxidase [Glycine max] 43 AAA19113 458119 acyl-CoA oxidase 44 AAA18595 495475 peroxisomal fatty acyl-coA oxidase 45 XP_540441 73965013 PREDICTED: similar to acyl-Coenzyme A oxidase isoform a [Canis familiaris] 46 NP_001 . . . 54400372 hypothetical protein LOC449662 [Danio rerio] 47 NP_001 . . . 78369480 acyl-Coenzyme A oxidase 1, palmitoyl [Bos taurus] 48 NP_004026 30089972 acyl-Coenzyme A oxidase isoform a [Homo sapiens] 49 AAH08767 14250616 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 50 CAD97622 31873262 hypothetical protein [Homo sapiens] 51 I38095 2117541 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal - human 52 NP_001 . . . 55741614 acyl-Coenzyme A oxidase 1, palmitoyl [Gallus gallus] 53 CAA68660 2673 acyl-coenzyme A oxidase [Candida tropicalis] 54 Q8HYL8 34098564 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 55 B54942 1082171 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal splice form II - human 56 AAH10425 14714578 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 57 CAJ83302 89271321 acyl-Coenzyme A oxidase 2, branched chain [Xenopus tropicalis] 58 AAO15577 27462768 acyl-CoA oxidase type 2 [Phascolarctos cinereus] 59 XP_414406 50754485 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Gallus gallus] 60 AAH97101 67678187 Zgc: 92584 protein [Danio rerio] 61 AAH85743 55249685 Acyl-Coenzyme A oxidase 1, palmitoyl [Rattus norvegicus] 62 2DDHA 93279231 Chain A, Crystal Structure Of Acyl-Coa Oxidase Complexed With 3-Oh-Dodecanoate 63 Q9ZQP2 62286640 Putative acyl-coenzyme A oxidase 1.2, peroxisomal 64 AAH63727 39795632 MGC68531 protein [Xenopus laevis] 65 NP_001 . . . 62751552 MGC108278 protein [Xenopus tropicalis] 66 AAH68891 46250227 MGC83074 protein [Xenopus laevis] 67 AAW78689 58531948 peroxisomal acyl-CoA oxidase 1A [Lycopersicon esculentum] 68 2FONC 109157678 Chain C, X-Ray Crystal Structure Of Leacx1, An Acyl-Coa Oxidase From Lycopersicon Esculentum (Tomato) 69 ABE83706 92875906 acyl-CoA oxidase [Medicago truncatula] 70 XP_511690 55645995 PREDICTED: similar to hypothetical protein [Pan troglodytes] 71 AAW78691 58531952 peroxisomal acyl-CoA oxidase 1A [Lycopersicon cheesmaniae] 72 AAL01887 15553478 acyl-CoA oxidase [Glycine max] 73 Q9Z1N0 17367267 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 74 BAA86870 6429156 peroxisomal acyl-CoA oxidase [Mus musculus] 75 Q9R0H0 51338830 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 76 BAF18456 113594582 Os06g0103500 [Oryza sativa (japonica cultivar-group)] 77 AAI08648 83318302 Unknown (protein for MGC: 131363) [Xenopus laevis] 78 AAB62926 2253380 peroxisomal acyl-CoA oxidase [Mus musculus] 79 NP_056544 66793429 acyl-Coenzyme A oxidase 1, palmitoyl [Mus musculus] 80 XP_320717 58394415 ENSANGP00000020118 [Anopheles gambiae str. PEST] 81 AAH21339 18204156 Acox2 protein [Mus musculus] 82 BAE25794 74188249 unnamed protein product [Mus musculus] 83 Q9QXD1 17367045 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 84 XP_320718 58394417 ENSANGP00000020032 [Anopheles gambiae str. PEST] 85 EAT39640 108875415 acyl-CoA oxidase [Aedes aegypti] 86 NP_665713 21955130 acyl-Coenzyme A oxidase 2, branched chain [Rattus norvegicus] 87 XP_625136 66556360 PREDICTED: similar to CG5009-PA, partial [Apis mellifera] 88 CAE63479 39592402 Hypothetical protein CBG07946 [Caenorhabditis briggsae] 89 XP_879996 76648809 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 6 [Bos taurus] 90 BAC26167 26324826 unnamed protein product [Mus musculus] 91 XP_592892 76648807 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 1 [Bos taurus] 92 AAH47700 28839704 Acyl-Coenzyme A oxidase 2, branched chain [Homo sapiens] 93 XP_541826 73985284 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Canis familiaris] 94 XP_782824 72015136 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 95 EAR95568 89297580 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 96 CAE63477 39592400 Hypothetical protein CBG07944 [Caenorhabditis briggsae] 97 NP_510603 17568313 F59F4.1 [Caenorhabditis elegans] 98 EAT48205 108883980 acyl-CoA oxidase [Aedes aegypti] 99 XP_785210 72015040 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 100 O02767 17366131 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 101 AAW78690 58531950 peroxisomal acyl-CoA oxidase 1B [Lycopersicon esculentum] 102 CAE63480 39592403 Hypothetical protein CBG07947 [Caenorhabditis briggsae] 103 EAS31618 90301987 hypothetical protein CIMG_07097 [Coccidioides immitis RS] 104 CAE63476 39592399 Hypothetical protein CBG07943 [Caenorhabditis briggsae] 105 XP_973660 91082769 PREDICTED: similar to CG5009-PA [Tribolium castaneum] 106 CAE69737 39596101 Hypothetical protein CBG16008 [Caenorhabditis briggsae] 107 NP_611264 24654595 CG5009-PA [Drosophila melanogaster] 108 AAF73843 8163758 acyl-CoA oxidase ACX3 [Arabidopsis thaliana] 109 Q9LLH9 62286634 Acyl-coenzyme A oxidase 3, peroxisomal precursor (AOX 3) (Medium-chain acyl-CoA oxidase) (AtCX3) 110 EAL26359 54636956 GA18591-PA [Drosophila pseudoobscura] 111 AAH54727 68262425 Acox1 protein [Mus musculus] 112 XP_783450 72111190 PREDICTED: similar to acyl-Coenzyme A oxidase 1, palmitoyl [Strongylocentrotus purpuratus] 113 XP_758355 71010163 hypothetical protein UM02208.1 [Ustilago maydis 521] 114 AAL28144 16767852 GH01266p [Drosophila melanogaster] 115 NP_001 . . . 71983346 F08A8.1a [Caenorhabditis elegans] 116 EAS04704 89306716 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 117 NP_493263 17506533 F08A8.3 [Caenorhabditis elegans] 118 BAC35782 74197284 unnamed protein product [Mus musculus] 119 XP_635946 66804335 hypothetical protein DDBDRAFT_0188674 [Dictyostelium discoideum AX4] 120 NP_493264 17506535 F08A8.4 [Caenorhabditis elegans] 121 CAJ58493 85539740 Hypothetical protein F08A8.1c [Caenorhabditis elegans] 122 NP_001 . . . 71983355 F08A8.1b [Caenorhabditis elegans] 123 EAS02716 89304728 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 124 XP_879827 76648805 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 5 [Bos taurus] 125 CAH91864 55730283 hypothetical protein [Pongo pygmaeus] 126 NP_499119 17552648 C48B4.1 [Caenorhabditis elegans] 127 ZP_012 . . . 90587482 Acyl-CoA oxidase-like:Acyl-CoA dehydrogenase-like:Acyl- CoA dehydrogenase, central region [Flavobacterium johnsoniae UW101] 128 BAE55537 83765394 unnamed protein product [Aspergillus oryzae] 129 XP_637610 66807775 hypothetical protein DDBDRAFT_0187085 [Dictyostelium discoideum AX4] 130 BAF19505 113595631 Os06g0354500 [Oryza sativa (japonica cultivar-group)] 131 EAT87650 111066530 hypothetical protein SNOG_05259 [Phaeosphaeria nodorum SN15] 132 CAE65171 39590798 Hypothetical protein CBG10043 [Caenorhabditis briggsae] hypothetical protein DDBDRAFT_0188084 [Dictyostelium 133 XP_636605 66805767 discoideum AX4] 134 XP_664356 67541164 hypothetical protein AN6752.2 [Aspergillus nidulans FGSC A4] 135 EAR99631 89301643 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 136 NP_493262 17506531 F08A8.2 [Caenorhabditis elegans] 137 XP_382463 46110811 hypothetical protein FG02287.1 [Gibberella zeae PH-1] 138 AAL39944 17862934 SD03592p [Drosophila melanogaster] 139 NP_523802 17647123 acyl-Coenzyme A oxidase at 57D proximal CG9707-PA [Drosophila melanogaster] 140 XP_587157 76620513 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Bos taurus] 141 EAR98581 89300593 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 142 Q9LMI7 62286635 Putative acyl-coenzyme A oxidase 3.2, peroxisomal precursor 143 O65201 62286588 Acyl-coenzyme A oxidase 2, peroxisomal precursor (AOX 2) (Long-chain acyl-CoA oxidase) (AtCX2) 144 AAC13497 3044212 acyl-CoA oxidase [Arabidopsis thaliana] 145 CAH91120 55728762 hypothetical protein [Pongo pygmaeus] 146 EAR89261 89291273 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 147 XP_646395 66826081 hypothetical protein DDBDRAFT_0216722 [Dictyostelium discoideum AX4] 148 O64894 62286587 Acyl-coenzyme A oxidase, peroxisomal precursor (AOX) (Long-chain acyl-CoA oxidase) 149 NP_998312 47085909 pristanoyl acyl-Coenzyme A oxidase 3 [Danio rerio] 150 O15254 17366151 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 151 AAL48010 17944231 LD22081p [Drosophila melanogaster] 152 ZP_012 . . . 89890388 acyl-CoA oxidase [Flavobacteria bacterium BBFL7] 153 XP_758113 71007469 hypothetical protein UM01966.1 [Ustilago maydis 521] 154 EAL26647 54637244 GA21980-PA [Drosophila pseudoobscura] 155 NP_109646 34328334 acyl-Coenzyme A oxidase 3, pristanoyl [Mus musculus] 156 YP_290295 72162638 hypothetical protein Tfu_2239 [Thermobifida fusca YX] 157 AAH44725 28703869 Acox3 protein [Mus musculus] 158 NP_445791 16758056 acyl-Coenzyme A oxidase 3, pristanoyl [Rattus norvegicus] 159 YP_445820 83816544 acyl-coenzyme A oxidase I, putative [Salinibacter ruber DSM 13855] 160 AAB67883 1575556 acyl-CoA oxidase homolog [Phalaenopsis sp. ‘True Lady’] 161 AAR00586 37699750 acyl-CoA oxidase [Phalaenopsis cv. ‘True Lady’] 162 XP_969513 91093755 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Tribolium castaneum] 163 XP_786025 72005645 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 164 YP_634556 108758121 putative acyl-CoA dehydrogenase [Myxococcus xanthus DK 1622] 165 EAT87454 111066334 hypothetical protein SNOG_05063 [Phaeosphaeria nodorum SN15] 166 Q9EPL9 17366740 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 167 XP_545908 73951751 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Canis familiaris] 168 EAL26648 54637245 GA21981-PA [Drosophila pseudoobscura] 169 BAC26136 26324764 unnamed protein product [Mus musculus] 170 CAA04688 2370232 putative acyl-CoA oxidase [Hordeum vulgare subsp. vulgare] 171 BAE42553 74143079 unnamed protein product [Mus musculus] 172 YP_702012 111019040 acyl-CoA oxidase [Rhodococcus sp. RHA1] 173 XP_786081 72005647 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 174 NP_001 . . . 79332306 ACX2 (ACYL-COA OXIDASE 2); acyl-CoA oxidase [Arabidopsis thaliana] 175 NP_572371 24640268 CG4586-PA [Drosophila melanogaster] 176 NP_961035 41408199 hypothetical protein MAP2101 [Mycobacterium avium subsp. paratuberculosis K-10] 177 EAL32329 54643586 GA18278-PA [Drosophila pseudoobscura] 178 ZP_012 . . . 90205075 Acyl-CoA oxidase-like:Acyl-CoA dehydrogenase-like:Acyl- CoA dehydrogenase, central region [Mycobacterium vanbaalenii PYR-1] 179 ABA94670 108864573 Acyl-coenzyme A oxidase 2, peroxisomal precursor, putative, expressed [Oryza sativa (japonica cultivar-group)] 180 BAE60166 83770031 unnamed protein product [Aspergillus oryzae] 181 ZP_011 . . . 89339305 Acyl-CoA oxidase-like:Acyl-CoA dehydrogenase-like:Acyl- CoA dehydrogenase, central region [Mycobacterium flavescens PYR-GCK] 182 NP_609027 45550141 CG9527-PA [Drosophila melanogaster] 183 EAT48670 108884445 acyl-CoA oxidase [Aedes aegypti] 184 ZP_012 . . . 92908524 Acyl-CoA oxidase-like:Acyl-CoA dehydrogenase-like:Acyl- CoA dehydrogenase, central region [Mycobacterium sp. JLS] 185 AAH17053 16877606 ACOX3 protein [Homo sapiens] 186 ZP_004 . . . 66967405 Acyl-CoA oxidase:Acyl-CoA dehydrogenase, C-terminal [Arthrobacter sp. FB24] 187 NP_508036 17560134 F25C8.1 [Caenorhabditis elegans] 188 ZP_011 . . . 88854811 acyl-CoA oxidase [marine actinobacterium PHSC20C1] 189 AAL48950 17945806 RE34879p [Drosophila melanogaster] 190 YP_640598 108800401 acyl-CoA dehydrogenase-like protein [Mycobacterium sp. MCS] 191 NP_724181 24585195 CG17544-PC, isoform C [Drosophila melanogaster] 192 AAL14003 16186117 SD05719p [Drosophila melanogaster] 193 XP_642669 66817952 hypothetical protein DDBDRAFT_0169270 [Dictyostelium discoideum AX4] 194 CAA96917 1945301 POX1 [Saccharomyces cerevisiae] 195 NP_500943 17540842 F58F9.7 [Caenorhabditis elegans] 196 XP_664369 67541190 hypothetical protein AN6765.2 [Aspergillus nidulans FGSC A4]

TABLE 35 Examples of pox1a polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_504703 50554589 YlPOX1 [Yarrowia lipolytica]. 2 O74934 59799072 Acyl-coenzyme A oxidase 1 (Acyl-CoA oxidase 1) 3 O74936 59799074 Acyl-coenzyme A oxidase 3 (Acyl-CoA oxidase 3) 4 XP_502199 50549457 YlPOX5 [Yarrowia lipolytica] 5 O74935 59799073 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) 6 XP_503632 50552444 hypothetical protein [Yarrowia lipolytica] 7 XP_504475 50554133 YlPOX4 [Yarrowia lipolytica] 8 XP_760471 71020481 hypothetical protein UM04324.1 [Ustilago maydis 521] 9 XP_571768 58269224 Acyl-coenzyme A oxidase I [Cryptococcus neoformans var. neoformans JEC21] 10 XP_717995 68475844 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 11 P08790 17373073 Acyl-coenzyme A oxidase 5 (Acyl-CoA oxidase 5) (PXP-5) 12 OXCKX5 66061 acyl-CoA oxidase (EC 1.3.3.6) POX5, peroxisomal - yeast (Candida tropicalis ) 13 Q9Y7B1 60391213 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 14 Q6BRD5 59799027 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 15 XP_457726 50417804 hypothetical protein DEHA0C01155g [Debaryomyces hansenii CBS767] 16 AAA34362 170912 acyl-coenzyme A oxidase II precursor 17 XP_459232 50421365 hypothetical protein DEHA0D18667g [Debaryomyces hansenii CBS767] 18 BAA83482 5763520 acyl-CoA oxidase [Candida tropicalis] 19 BAA83483 5763522 acyl-CoA oxidase [Candida tropicalis] 20 OXCKPM 66060 acyl-CoA oxidase (EC 1.3.3.6) PXP4, peroxisomal - yeast (Candida maltosa ) 21 AAA34361 170910 PXP-2 protein 22 XP_721610 68468582 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 23 1306283A 225549 oxidase, fatty acyl 24 P06598 17373023 Acyl-coenzyme A oxidase 4 (Acyl-CoA oxidase 4) (PXP-4) (Peroxisomal fatty acyl-CoA oxidase) 25 XP_721613 68468588 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 26 Q00468 2498206 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) (AOX 2) 27 Q6CKK7 59799028 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 28 Q6FY63 59799032 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 29 Q756A9 60391210 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 30 NP_011310 6321233 Fatty-acyl coenzyme A oxidase, involved in the fatty acid beta- oxidation pathway; localized to the peroxisomal matrix; Pox1p [Saccharomyces cerevisiae] 31 AAA34891 172217 acyl-coenzyme A oxidase 32 O65202 62286589 Acyl-coenzyme A oxidase 1, peroxisomal (AOX 1) (Long-chain acyl-CoA oxidase) (AtCX1) 33 1W07B 58177067 Chain B, Arabidopsis Thaliana Acyl-Coa Oxidase 1 34 H71434 7488059 probable apetala2 domain TINY - Arabidopsis thaliana 35 NP_567513 18414744 ACX1 (ACYL-COA OXIDASE 1) [Arabidopsis thaliana] 36 Q9ZQP2 62286640 Putative acyl-coenzyme A oxidase 1.2, peroxisomal 37 AAL01888 15553480 acyl-CoA oxidase [Glycine max] 38 CAA68660 2673 acyl-coenzyme A oxidase [Candida tropicalis] 39 AAL01887 15553478 acyl-CoA oxidase [Glycine max] 40 BAF18456 113594582 Os06g0103500 [Oryza sativa (japonica cultivar-group)] 41 NP_001 . . . 55741614 acyl-Coenzyme A oxidase 1, palmitoyl [Gallus gallus] 42 AAH68891 46250227 MGC83074 protein [Xenopus laevis] 43 2FONC 109157678 Chain C, X-Ray Crystal Structure Of Leacx1, An Acyl-Coa Oxidase From Lycopersicon Esculentum (Tomato) 44 NP_001 . . . 54400372 hypothetical protein LOC449662 [Danio rerio] 45 AAW78689 58531948 peroxisomal acyl-CoA oxidase 1A [Lycopersicon esculentum] 46 XP_001 . . . 109118312 PREDICTED: similar to acyl-Coenzyme A oxidase isoform b [Macaca mulatta] 47 AAW78691 58531952 peroxisomal acyl-CoA oxidase 1A [Lycopersicon cheesmaniae] 48 Q5RC19 62286600 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 49 ABE83706 92875906 acyl-CoA oxidase [Medicago truncatula] 50 NP_009223 30089974 acyl-Coenzyme A oxidase isoform b [Homo sapiens] 51 Q15067 17380467 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) (Straight-chain acyl-CoA oxidase) (SCOX) 52 AAA18595 495475 peroxisomal fatty acyl-coA oxidase 53 AAA19113 458119 acyl-CoA oxidase 54 NP_001 . . . 78369480 acyl-Coenzyme A oxidase 1, palmitoyl [Bos taurus] 55 AAH63727 39795632 MGC68531 protein [Xenopus laevis] 56 AAB30019 7689914 peroxisomal acyl-coenzyme A oxidase [Homo sapiens] 57 NP_001 . . . 62751552 MGC108278 protein [Xenopus tropicalis] 58 BAA86870 6429156 peroxisomal acyl-CoA oxidase [Mus musculus] 59 Q9R0H0 51338830 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 60 AAH85743 55249685 Acyl-Coenzyme A oxidase 1, palmitoyl [Rattus norvegicus] 61 AAB62926 2253380 peroxisomal acyl-CoA oxidase [Mus musculus] 62 CAH90691 55727877 hypothetical protein [Pongo pygmaeus] 63 CAJ83302 89271321 acyl-Coenzyme A oxidase 2, branched chain [Xenopus tropicalis] 64 AAH97101 67678187 Zgc:92584 protein [Danio rerio] 65 NP_004026 30089972 acyl-Coenzyme A oxidase isoform a [Homo sapiens] 66 AAH08767 14250616 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 67 CAD97622 31873262 hypothetical protein [Homo sapiens] 68 B54942 1082171 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal splice form II - human 69 2DDHA 93279231 Chain A, Crystal Structure Of Acyl-Coa Oxidase Complexed With 3-Oh-Dodecanoate 70 XP_540441 73965013 PREDICTED: similar to acyl-Coenzyme A oxidase isoform a [Canis familiaris] 71 I38095 2117541 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal - human 72 AAH10425 14714578 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 73 NP_056544 66793429 acyl-Coenzyme A oxidase 1, palmitoyl [Mus musculus] 74 AAI08648 83318302 Unknown (protein for MGC:131363) [Xenopus laevis] 75 Q9Z1N0 17367267 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 76 XP_414406 50754485 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Gallus gallus] 77 Q8HYL8 34098564 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 78 XP_511690 55645995 PREDICTED: similar to hypothetical protein [Pan troglodytes] 79 AAO15577 27462768 acyl-CoA oxidase type 2 [Phascolarctos cinereus] 80 XP_625136 66556360 PREDICTED: similar to CG5009-PA, partial [Apis mellifera] 81 XP_320717 58394415 ENSANGP00000020118 [Anopheles gambiae str. PEST] 82 EAT39640 108875415 acyl-CoA oxidase [Aedes aegypti] 83 BAC26167 26324826 unnamed protein product [Mus musculus] 84 EAS31618 90301987 hypothetical protein CIMG_07097 [Coccidioides immitis RS] 85 XP_382463 46110811 hypothetical protein FG02287.1 [Gibberella zeae PH-1] 86 XP_320718 58394417 ENSANGP00000020032 [Anopheles gambiae str. PEST] 87 CAE63479 39592402 Hypothetical protein CBG07946 [Caenorhabditis briggsae] 88 AAW78690 58531950 peroxisomal acyl-CoA oxidase 1B [Lycopersicon esculentum] 89 CAE69737 39596101 Hypothetical protein CBG16008 [Caenorhabditis briggsae] 90 AAH21339 18204156 Acox2 protein [Mus musculus] 91 BAE25794 74188249 unnamed protein product [Mus musculus] 92 EAR95568 89297580 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 93 BAE55537 83765394 unnamed protein product [Aspergillus oryzae] 94 Q9QXD1 17367045 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 95 NP_510603 17568313 F59F4.1 [Caenorhabditis elegans] 96 XP_785210 72015040 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 97 XP_973660 91082769 PREDICTED: similar to CG5009-PA [Tribolium castaneum] 98 XP_782824 72015136 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 99 AAH47700 28839704 Acyl-Coenzyme A oxidase 2, branched chain [Homo sapiens] 100 CAE63480 39592403 Hypothetical protein CBG07947 [Caenorhabditis briggsae] 101 O02767 17366131 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 102 NP_665713 21955130 acyl-Coenzyme A oxidase 2, branched chain [Rattus norvegicus] 103 XP_664356 67541164 hypothetical protein AN6752.2 [Aspergillus nidulans FGSC A4] 104 EAT48205 108883980 acyl-CoA oxidase [Aedes aegypti] 105 CAE63477 39592400 Hypothetical protein CBG07944 [Caenorhabditis briggsae] 106 XP_758355 71010163 hypothetical protein UM02208.1 [Ustilago maydis 521] 107 CAE65171 39590798 Hypothetical protein CBG10043 [Caenorhabditis briggsae] 108 XP_637610 66807775 hypothetical protein DDBDRAFT_0187085 [Dictyostelium discoideum AX4] 109 NP_499119 17552648 C48B4.1 [Caenorhabditis elegans] 110 XP_635946 66804335 hypothetical protein DDBDRAFT_0188674 [Dictyostelium discoideum AX4] 111 XP_592892 76648807 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 1 [Bos taurus] 112 XP_879996 76648809 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 6 [Bos taurus] 113 XP_783450 72111190 PREDICTED: similar to acyl-Coenzyme A oxidase 1, palmitoyl [Strongylocentrotus purpuratus] 114 XP_541826 73985284 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Canis familiaris] 115 EAR89261 89291273 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 116 AAH54727 68262425 Acox1 protein [Mus musculus] 117 BAC35782 74197284 unnamed protein product [Mus musculus] 118 NP_493263 17506533 F08A8.3 [Caenorhabditis elegans] 119 AAF14635 6503198 acyl-CoA oxidase [Petroselinum crispum] 120 CAE63476 39592399 Hypothetical protein CBG07943 [Caenorhabditis briggsae] 121 AAL39944 17862934 SD03592p [Drosophila melanogaster] 122 NP_523802 17647123 acyl-Coenzyme A oxidase at 57D proximal CG9707-PA [Drosophila melanogaster] 123 ZP_012 . . . 90587482 Acyl-CoA oxidase-like:Acyl-CoA dehydrogenase-like:Acyl- CoA dehydrogenase, central region [Flavobacterium johnsoniae UW101] 124 AAR00586 37699750 acyl-CoA oxidase [Phalaenopsis cv. ‘True Lady’] 125 XP_758113 71007469 hypothetical protein UM01966.1 [Ustilago maydis 521] 126 AAB67883 1575556 acyl-CoA oxidase homolog [Phalaenopsis sp. ‘True Lady’] 127 NP_001 . . . 71983346 F08A8.1a [Caenorhabditis elegans] 128 NP_001 . . . 71983355 F08A8.1b [Caenorhabditis elegans] 129 CAJ58493 85539740 Hypothetical protein F08A8.1c [Caenorhabditis elegans] 130 AAC13497 3044212 acyl-CoA oxidase [Arabidopsis thaliana] 131 O65201 62286588 Acyl-coenzyme A oxidase 2, peroxisomal precursor (AOX 2) (Long-chain acyl-CoA oxidase) (AtCX2) 132 EAS04704 89306716 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 133 EAL26359 54636956 GA18591-PA [Drosophila pseudoobscura] 134 XP_636605 66805767 hypothetical protein DDBDRAFT_0188084 [Dictyostelium discoideum AX4] 135 O64894 62286587 Acyl-coenzyme A oxidase, peroxisomal precursor (AOX) (Long-chain acyl-CoA oxidase) 136 NP_611264 24654595 CG5009-PA [Drosophila melanogaster] 137 NP_493264 17506535 F08A8.4 [Caenorhabditis elegans] 138 AAH44725 28703869 Acox3 protein [Mus musculus] 139 NP_109646 34328334 acyl-Coenzyme A oxidase 3, pristanoyl [Mus musculus] 140 XP_786025 72005645 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 141 XP_646395 66826081 hypothetical protein DDBDRAFT_0216722 [Dictyostelium discoideum AX4] 142 EAT87650 111066530 hypothetical protein SNOG_05259 [Phaeosphaeria nodorum SN15] 143 NP_998312 47085909 pristanoyl acyl-Coenzyme A oxidase 3 [Danio rerio] 144 YP_634556 108758121 putative acyl-CoA dehydrogenase [Myxococcus xanthus DK 1622] 145 AAL28144 16767852 GH01266p [Drosophila melanogaster] 146 YP_445820 83816544 acyl-coenzyme A oxidase I, putative [Salinibacter ruber DSM 13855] 147 EAL26647 54637244 GA21980-PA [Drosophila pseudoobscura] 148 XP_786081 72005647 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 149 EAR99631 89301643 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 150 NP_493262 17506531 F08A8.2 [Caenorhabditis elegans] 151 CAH91864 55730283 hypothetical protein [Pongo pygmaeus] 152 Q9LLH9 62286634 Acyl-coenzyme A oxidase 3, peroxisomal precursor (AOX 3) (Medium-chain acyl-CoA oxidase) (AtCX3) 153 AAF73843 8163758 acyl-CoA oxidase ACX3 [Arabidopsis thaliana] 154 XP_969513 91093755 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Tribolium castaneum] 155 O15254 17366151 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 156 Q9EPL9 17366740 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 157 NP_445791 16758056 acyl-Coenzyme A oxidase 3, pristanoyl [Rattus norvegicus] 158 ABA94670 108864573 Acyl-coenzyme A oxidase 2, peroxisomal precursor, putative, expressed [Oryza sativa (japonica cultivar-group)] 159 EAT87454 111066334 hypothetical protein SNOG_05063 [Phaeosphaeria nodorum SN15] 160 EAL26648 54637245 GA21981-PA [Drosophila pseudoobscura] 161 EAR98581 89300593 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 162 BAE42553 74143079 unnamed protein product [Mus musculus] 163 BAC26136 26324764 unnamed protein product [Mus musculus] 164 XP_879827 76648805 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 5 [Bos taurus] 165 XP_587157 76620513 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Bos taurus] 166 YP_290295 72162638 hypothetical protein Tfu_2239 [Thermobifida fusca YX] 167 CAH91120 55728762 hypothetical protein [Pongo pygmaeus] 168 EAS02716 89304728 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 169 AAL48010 17944231 LD22081p [Drosophila melanogaster] 170 XP_545908 73951751 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Canis familiaris] 171 ZP_011 . . . 88854811 acyl-CoA oxidase [marine actinobacterium PHSC20C1] 172 NP_001 . . . 79332306 ACX2 (ACYL-COA OXIDASE 2); acyl-CoA oxidase [Arabidopsis thaliana] 173 BAF19505 113595631 Os06g0354500 [Oryza sativa (japonica cultivar-group)] 174 EAL32329 54643586 GA18278-PA [Drosophila pseudoobscura] 175 CAA04688 2370232 putative acyl-CoA oxidase [Hordeum vulgare subsp. vulgare] 176 ZP_012 . . . 89890388 acyl-CoA oxidase [Flavobacteria bacterium BBFL7] 177 Q9LMI7 62286635 Putative acyl-coenzyme A oxidase 3.2, peroxisomal precursor 178 XP_395486 66564429 PREDICTED: similar to CG9527-PA [Apis mellifera] 179 YP_702012 111019040 acyl-CoA oxidase [Rhodococcus sp. RHA1] 180 ZP_012 . . . 90205075 Acyl-CoA oxidase-like:Acyl-CoA dehydrogenase-like:Acyl- CoA dehydrogenase, central region [Mycobacterium vanbaalenii PYR-1] 181 AAL48950 17945806 RE34879p [Drosophila melanogaster] 182 NP_609027 45550141 CG9527-PA [Drosophila melanogaster] 183 ZP_006 . . . 71369913 Acyl-CoA oxidase:Acyl-CoA dehydrogenase, C-terminal:Acyl- CoA dehydrogenase, central region:Acyl-CoA dehydrogenase, N-terminal [Nocardioides sp. JS614] 184 ZP_012 . . . 92908524 Acyl-CoA oxidase-like:Acyl-CoA dehydrogenase-like:Acyl- CoA dehydrogenase, central region [Mycobacterium sp. JLS] 185 EAT48670 108884445 acyl-CoA oxidase [Aedes aegypti] 186 XP_317446 58390030 ENSANGP00000011863 [Anopheles gambiae str. PEST] 187 EAS36523 90306892 hypothetical protein CIMG_01877 [Coccidioides immitis RS] 188 NP_572371 24640268 CG4586-PA [Drosophila melanogaster] 189 XP_642669 66817952 hypothetical protein DDBDRAFT_0169270 [Dictyostelium discoideum AX4] 190 CAA96917 1945301 POX1 [Saccharomyces cerevisiae] 191 ZP_011 . . . 89339305 Acyl-CoA oxidase-like:Acyl-CoA dehydrogenase-like:Acyl- CoA dehydrogenase, central region [Mycobacterium flavescens PYR-GCK] 192 ZP_003 . . . 62426247 COG1960: Acyl-CoA dehydrogenases [Brevibacterium linens BL2] 193 YP_640598 108800401 acyl-CoA dehydrogenase-like protein [Mycobacterium sp. MCS] 194 NP_724181 24585195 CG17544-PC, isoform C [Drosophila melanogaster] 195 AAL14003 16186117 SD05719p [Drosophila melanogaster] 196 AAH17053 16877606 ACOX3 protein [Homo sapiens] 197 EAL32912 54644170 GA21857-PA [Drosophila pseudoobscura]

TABLE 36 Examples of pox4 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_504475.1 50554133 YlPOX4 [Yarrowia lipolytica] 2 O74935 59799073 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) 3 O74936 59799074 Acyl-coenzyme A oxidase 3 (Acyl-CoA oxidase 3) 4 XP_502199 50549457 YlPOX5 [Yarrowia lipolytica] 5 O74934 59799072 Acyl-coenzyme A oxidase 1 (Acyl-CoA oxidase 1) 6 XP_504703 50554589 YlPOX1 [Yarrowia lipolytica] 7 XP_503632 50552444 hypothetical protein [Yarrowia lipolytica] 8 XP_760471 71020481 hypothetical protein UM04324.1 [Ustilago maydis 521] 9 XP_571768 58269224 Acyl-coenzyme A oxidase I [Cryptococcus neoformans var. neoformans JEC21] 10 XP_717995 68475844 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 11 BAA83482 5763520 acyl-CoA oxidase [Candida tropicalis] 12 BAA83483 5763522 acyl-CoA oxidase [Candida tropicalis] 13 OXCKPM 66060 acyl-CoA oxidase (EC 1.3.3.6) PXP4, peroxisomal-yeast (Candida maltosa) 14 XP_721610 68468582 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 15 AAA34362 170912 acyl-coenzyme A oxidase II precursor 16 P08790 17373073 Acyl-coenzyme A oxidase 5 (Acyl-CoA oxidase 5) (PXP-5) 17 OXCKX5 66061 acyl-CoA oxidase (EC 1.3.3.6) POX5, peroxisomal-yeast (Candida tropicalis) 18 Q6BRD5 59799027 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 19 XP_457726 50417804 hypothetical protein DEHA0C01155g [Debaryomyces hansenii CBS767] 20 P06598 17373023 Acyl-coenzyme A oxidase 4 (Acyl-CoA oxidase 4) (PXP-4) (Peroxisomal fatty acyl-CoA oxidase) 21 1306283A 225549 oxidase, fatty acyl 22 AAA34361 170910 PXP-2 protein 23 XP_459232 50421365 hypothetical protein DEHA0D18667g [Debaryomyces hansenii CBS767] 24 Q9Y7B1 60391213 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 25 XP_721613 68468588 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 26 Q00468 2498206 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) (AOX 2) 27 Q6FY63 59799032 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 28 NP_011310 6321233 Fatty-acyl coenzyme A oxidase, involved in the fatty acid beta- oxidation pathway; localized to the peroxisomal matrix; Pox1p [Saccharomyces cerevisiae] 29 AAA34891 172217 acyl-coenzyme A oxidase 30 Q6CKK7 59799028 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 31 Q756A9 60391210 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 32 O65202 62286589 Acyl-coenzyme A oxidase 1, peroxisomal (AOX 1) (Long-chain acyl-CoA oxidase) (AtCX1) 33 CAA68660 2673 acyl-coenzyme A oxidase [Candida tropicalis] 34 1W07B 58177067 Chain B, Arabidopsis Thaliana Acyl-Coa Oxidase 1 35 AAL01888 15553480 acyl-CoA oxidase [Glycine max] 36 H71434 7488059 probable apetala2 domain TINY-Arabidopsis thaliana 37 NP_567513 18414744 ACX1 (ACYL-COA OXIDASE 1) [Arabidopsis thaliana] 38 AAH63727 39795632 MGC68531 protein [Xenopus laevis] 39 AAL01887 15553478 acyl-CoA oxidase [Glycine max] 40 AAI08648 83318302 Unknown (protein for MGC: 131363) [Xenopus laevis] 41 BAF18456 113594582 Os06g0103500 [Oryza sativa (japonica cultivar-group)] 42 Q9ZQP2 62286640 Putative acyl-coenzyme A oxidase 1.2, peroxisomal 43 NP_001 . . . 78369480 acyl-Coenzyme A oxidase 1, palmitoyl [Bos taurus] 44 ABE83706 92875906 acyl-CoA oxidase [Medicago truncatula] 45 NP_001 . . . 62751552 MGC108278 protein [Xenopus tropicalis] 46 NP_001 . . . 55741614 acyl-Coenzyme A oxidase 1, palmitoyl [Gallus gallus] 47 AAW78689 58531948 peroxisomal acyl-CoA oxidase 1A [Lycopersicon esculentum] 48 2FONC 109157678 Chain C, X-Ray Crystal Structure Of Leacx1, An Acyl-Coa Oxidase From Lycopersicon Esculentum (Tomato) 49 NP_001 . . . 54400372 hypothetical protein LOC449662 [Danio rerio] 50 AAW78691 58531952 peroxisomal acyl-CoA oxidase 1A [Lycopersicon cheesmaniae] 51 Q5RC19 62286600 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 52 XP_001 . . . 109118312 PREDICTED: similar to acyl-Coenzyme A oxidase isoform b [Macaca mulatta] 53 Q15067 17380467 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) (Straight-chain acyl-CoA oxidase) (SCOX) 54 NP_009223 30089974 acyl-Coenzyme A oxidase isoform b [Homo sapiens] 55 AAO15577 27462768 acyl-CoA oxidase type 2 [Phascolarctos cinereus] 56 AAB30019 7689914 peroxisomal acyl-coenzyme A oxidase [Homo sapiens] 57 AAH97101 67678187 Zgc: 92584 protein [Danio rerio] 58 XP_414406 50754485 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Gallus gallus] 59 AAA19113 458119 acyl-CoA oxidase 60 AAA18595 495475 peroxisomal fatty acyl-coA oxidase 61 Q9Z1N0 17367267 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 62 Q8HYL8 34098564 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 63 AAH85743 55249685 Acyl-Coenzyme A oxidase 1, palmitoyl [Rattus norvegicus] 64 AAH10425 14714578 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 65 CAH90691 55727877 hypothetical protein [Pongo pygmaeus] 66 CAD97622 31873262 hypothetical protein [Homo sapiens] 67 AAH08767 14250616 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 68 NP_004026 30089972 acyl-Coenzyme A oxidase isoform a [Homo sapiens] 69 I38095 2117541 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal-human 70 BAA86870 6429156 peroxisomal acyl-CoA oxidase [Mus musculus] 71 CAJ83302 89271321 acyl-Coenzyme A oxidase 2, branched chain [Xenopus tropicalis] 72 Q9R0H0 51338830 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 73 AAB62926 2253380 peroxisomal acyl-CoA oxidase [Mus musculus] 74 B54942 1082171 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal splice form II- human 75 XP_320717 58394415 ENSANGP00000020118 [Anopheles gambiae str. PEST] 76 BAE25794 74188249 unnamed protein product [Mus musculus] 77 AAH21339 18204156 Acox2 protein [Mus musculus] 78 2DDHA 93279231 Chain A, Crystal Structure Of Acyl-Coa Oxidase Complexed With 3-Oh- Dodecanoate 79 Q9QXD1 17367045 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 80 AAH68891 46250227 MGC83074 protein [Xenopus laevis] 81 XP_540441 73965013 PREDICTED: similar to acyl-Coenzyme A oxidase isoform a [Canis familiaris] 82 XP_320718 58394417 ENSANGP00000020032 [Anopheles gambiae str. PEST] 83 XP_511690 55645995 PREDICTED: similar to hypothetical protein [Pan troglodytes] 84 NP_056544 66793429 acyl-Coenzyme A oxidase 1, palmitoyl [Mus musculus] 85 EAT39640 108875415 acyl-CoA oxidase [Aedes aegypti] 86 XP_783450 72111190 PREDICTED: similar to acyl-Coenzyme A oxidase 1, palmitoyl [Strongylocentrotus purpuratus] 87 NP_665713 21955130 acyl-Coenzyme A oxidase 2, branched chain [Rattus norvegicus] 88 XP_973660 91082769 PREDICTED: similar to CG5009-PA [Tribolium castaneum] 89 CAE63476 39592399 Hypothetical protein CBG07943 [Caenorhabditis briggsae] 90 XP_625136 66556360 PREDICTED: similar to CG5009-PA, partial [Apis mellifera] 91 XP_592892 76648807 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 1 [Bos taurus] 92 XP_879996 76648809 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 6 [Bos taurus] 93 AAW78690 58531950 peroxisomal acyl-CoA oxidase 1B [Lycopersicon esculentum] 94 AAH47700 28839704 Acyl-Coenzyme A oxidase 2, branched chain [Homo sapiens] 95 CAE69737 39596101 Hypothetical protein CBG16008 [Caenorhabditis briggsae] 96 NP_499119 17552648 C48B4.1 [Caenorhabditis elegans] 97 CAE65171 39590798 Hypothetical protein CBG10043 [Caenorhabditis briggsae] 98 XP_541826 73985284 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Canis familiaris] 99 NP_510603 17568313 F59F4.1 [Caenorhabditis elegans] 100 NP_001 . . . 71983346 F08A8.1a [Caenorhabditis elegans] 101 EAR95568 89297580 Acyl-CoA oxidase family protein [Tetrahymena thermophila] SB210] 102 NP_001 . . . 71983355 F08A8.1b [Caenorhabditis elegans] 103 CAJ58493 85539740 Hypothetical protein F08A8.1c [Caenorhabditis elegans] 104 EAL26359 54636956 GA18591-PA [Drosophila pseudoobscura] 105 BAC26167 26324826 unnamed protein product [Mus musculus] 106 CAE63479 39592402 Hypothetical protein CBG07946 [Caenorhabditis briggsae] 107 O02767 17366131 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 108 NP_611264 24654595 CG5009-PA [Drosophila melanogaster] 109 CAE63480 39592403 Hypothetical protein CBG07947 [Caenorhabditis briggsae] 110 XP_758355 71010163 hypothetical protein UM02208.1 [Ustilago maydis 521] 111 EAT48205 108883980 acyl-CoA oxidase [Aedes aegypti] 112 XP_782824 72015136 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 113 AAL28144 16767852 GH01266p [Drosophila melanogaster] 114 XP_785210 72015040 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 115 EAS31618 90301987 hypothetical protein CIMG_07097 [Coccidioides immitis RS] 116 XP_879827 76648805 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 5 [Bos taurus] 117 CAH91864 55730283 hypothetical protein [Pongo pygmaeus] 118 NP_493263 17506533 F08A8.3 [Caenorhabditis elegans] 119 Q9LLH9 62286634 Acyl-coenzyme A oxidase 3, peroxisomal precursor (AOX 3) (Medium-chain acyl-CoA oxidase) (AtCX3) 120 AAF73843 8163758 acyl-CoA oxidase ACX3 [Arabidopsis thaliana] 121 AAL39944 17862934 SD03592p [Drosophila melanogaster] 122 NP_523802 17647123 acyl-Coenzyme A oxidase at 57D proximal CG9707-PA [Drosophila melanogaster] 123 AAH54727 68262425 Acox1 protein [Mus musculus] 124 BAC35782 74197284 unnamed protein product [Mus musculus] 125 BAE55537 83765394 unnamed protein product [Aspergillus oryzae] 126 CAE63477 39592400 Hypothetical protein CBG07944 [Caenorhabditis briggsae] 127 XP_636605 66805767 hypothetical protein DDBDRAFT_0188084 [Dictyostelium discoideum AX4] 128 NP_493264 17506535 F08A8.4 [Caenorhabditis elegans] 129 NP_493262 17506531 F08A8.2 [Caenorhabditis elegans] 130 EAL26648 54637245 GA21981-PA [Drosophila pseudoobscura] 131 AAF14635 6503198 acyl-CoA oxidase [Petroselinum crispum] 132 XP_664356 67541164 hypothetical protein AN6752.2 [Aspergillus nidulans FGSC A4] 133 EAL26647 54637244 GA21980-PA [Drosophila pseudoobscura] 134 AAL48010 17944231 LD22081p [Drosophila melanogaster] 135 ZP_012 . . . 90587482 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Flavobacterium johnsoniae UW101] 136 CAA04688 2370232 putative acyl-CoA oxidase [Hordeum vulgare subsp. vulgare] 137 Q9LMI7 62286635 Putative acyl-coenzyme A oxidase 3.2, peroxisomal precursor 138 EAR89261 89291273 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 139 YP_290295 72162638 hypothetical protein Tfu_2239 [Thermobifida fusca YX] 140 XP_758113 71007469 hypothetical protein UM01966.1 [Ustilago maydis 521] 141 BAF19505 113595631 Os06g0354500 [Oryza sativa (japonica cultivar-group)] 142 XP_317446 58390030 ENSANGP00000011863 [Anopheles gambiae str. PEST] 143 XP_635946 66804335 hypothetical protein DDBDRAFT_0188674 [Dictyostelium discoideum AX4] 144 XP_382463 46110811 hypothetical protein FG02287.1 [Gibberella zeae PH-2] 145 YP_445820 83816544 acyl-coenzyme A oxidase I, putative [Salinibacter ruber DSM 13855] 146 EAR99631 89301643 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 147 EAT87650 111066530 hypothetical protein SNOG_05259 [Phaeosphaeria nodorum SN15] 148 O15254 17366151 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 149 NP_508036 17560134 F25C8.1 [Caenorhabditis elegans] 150 NP_998312 47085909 pristanoyl acyl-Coenzyme A oxidase 3 [Danio rerio] 151 EAL32329 54643586 GA18278-PA [Drosophila pseudoobscura] 152 EAS02716 89304728 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 153 EAT48670 108884445 acyl-CoA oxidase [Aedes aegypti] 154 CAH91120 55728762 hypothetical protein [Pongo pygmaeus] 155 EAS04704 89306716 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 156 ZP_012 . . . 89890388 acyl-CoA oxidase [Flavobacteria bacterium BBFL7] 157 O64894 62286587 Acyl-coenzyme A oxidase, peroxisomal precursor (AOX) (Long-chain acyl-CoA oxidase) 158 XP_969513 91093755 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Tribolium castaneum] 159 AAC13497 3044212 acyl-CoA oxidase [Arabidopsis thaliana] 160 O65201 62286588 Acyl-coenzyme A oxidase 2, peroxisomal precursor (AOX 2) (Long-chain acyl-CoA oxidase) (AtCX2) 161 XP_646395 66826081 hypothetical protein DDBDRAFT_0216722 [Dictyostelium discoideum AX4] 162 YP_634556 108758121 putative acyl-CoA dehydrogenase [Myxococcus xanthus DK 1622] 163 XP_587157 76620513 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Bos taurus] 164 EAR98581 89300593 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 165 XP_637610 66807775 hypothetical protein DDBDRAFT_0187085 [Dictyostelium discoideum AX4] 166 NP_572371 24640268 CG4586-PA [Drosophila melanogaster] 167 EAT87454 111066334 hypothetical protein SNOG_05063 [Phaeosphaeria nodorum SN15] 168 AAB67883 1575556 acyl-CoA oxidase homolog [Phalaenopsis sp. ‘True Lady’] 169 NP_961035 41408199 hypothetical protein MAP2101 [Mycobacterium avium subsp. paratuberculosis K-10] 170 ABA94670 108864573 Acyl-coenzyme A oxidase 2, peroxisomal precursor, putative, expressed [Oryza sativa (japonica cultivar-group)] 171 AAR00586 37699750 acyl-CoA oxidase [Phalaenopsis cv. ‘True Lady’] 172 XP_545908 73951751 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Canis familiaris] 173 YP_702012 111019040 acyl-CoA oxidase [Rhodococcus sp. RHA1] 174 ZP_012 . . . 90205075 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium vanbaalenii PYR-1] 175 EAS36523 90306892 hypothetical protein CIMG_01877 [Coccidioides immitis RS] 176 NP_001 . . . 79332306 ACX2 (ACYL-COA OXIDASE 2); acyl-CoA oxidase [Arabidopsis thaliana] 177 CAA68661 2674 44 kD translation product [Candida tropicalis] 178 ZP_012 . . . 92908524 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium sp. JLS] 179 XP_786025 72005645 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 180 CAA96917 1945301 POX1 [Saccharomyces cerevisiae] 181 BAE60166 83770031 unnamed protein product [Aspergillus oryzae] 182 ZP_011 . . . 89339305 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium flavescens PYR-GCK] 183 YP_640598 108800401 acyl-CoA dehydrogenase-like protein [Mycobacterium sp. MCS] 184 ZP_006 . . . 71369913 Acyl-CoA oxidase: Acyl-CoA dehydrogenase, C-terminal: Acyl- CoA dehydrogenase, central region: Acyl-CoA dehydrogenase, N-terminal [Nocardioides sp. JS614] 185 AAL14003 16186117 SD05719p [Drosophila melanogaster] 186 NP_724181 24585195 CG17544-PC, isoform C [Drosophila melanogaster] 187 AAH44725 28703869 Acox3 protein [Mus musculus] 188 NP_109646 34328334 acyl-Coenzyme A oxidase 3, pristanoyl [Mus musculus] 189 EAL33335 54644594 GA14550-PA [Drosophila pseudoobscura] 190 ZP_004 . . . 66967405 Acyl-CoA oxidase: Acyl-CoA dehydrogenase, C-terminal [Arthrobacter sp. FB24] 191 NP_445791 16758056 acyl-Coenzyme A oxidase 3, pristanoyl [Rattus norvegicus] 192 XP_786081 72005647 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 193 Q9EPL9 17366740 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 194 NP_609027 45550141 CG9527-PA [Drosophila melanogaster] 195 ZP_006 . . . 69287897 Acyl-CoA dehydrogenase, C-terminal: Acyl-CoA dehydrogenase, central region [Kineococcus radiotolerans SRS30216] 196 BAE47462 78483008 acyl-CoA oxidase [Arthrobacter ureafaciens]

TABLE 37 Examples of pox1 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_503632 50552444 hypothetical protein [Yarrowia lipolytica]. 2 O74935 59799073 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) 3 O74936 59799074 Acyl-coenzyme A oxidase 3 (Acyl-CoA oxidase 3) 4 XP_502199 50549457 YlPOX5 [Yarrowia lipolytica] 5 O74934 59799072 Acyl-coenzyme A oxidase 1 (Acyl-CoA oxidase 1) 6 XP_504703 50554589 YlPOX1 [Yarrowia lipolytica] 7 XP_504475 50554133 YlPOX4 [Yarrowia lipolytica] 8 XP_760471 71020481 hypothetical protein UM04324.1 [Ustilago maydis 521] 9 XP_571768 58269224 Acyl-coenzyme A oxidase I [Cryptococcus neoformans var. neoformans JEC21] 10 AAA34362 170912 acyl-coenzyme A oxidase II precursor 11 BAA83482 5763520 acyl-CoA oxidase [Candida tropicalis] 12 XP_717995 68475844 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 13 BAA83483 5763522 acyl-CoA oxidase [Candida tropicalis] 14 XP_721610 68468582 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 15 OXCKPM 66060 acyl-CoA oxidase (EC 1.3.3.6) PXP4, peroxisomal-yeast (Candida maltosa) 16 P08790 17373073 Acyl-coenzyme A oxidase 5 (Acyl-CoA oxidase 5) (PXP-5) 17 OXCKX5 66061 acyl-CoA oxidase (EC 1.3.3.6) POX5, peroxisomal-yeast (Candida tropicalis) 18 XP_721613 68468588 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 19 AAA34361 170910 PXP-2 protein 20 Q6BRD5 59799027 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 21 1306283A 225549 oxidase, fatty acyl 22 P06598 17373023 Acyl-coenzyme A oxidase 4 (Acyl-CoA oxidase 4) (PXP-4) (Peroxisomal fatty acyl-CoA oxidase) 23 XP_459232 50421365 hypothetical protein DEHA0D18667g [Debaryomyces hansenii CBS767] 24 Q00468 2498206 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) (AOX 2) 25 XP_457726 50417804 hypothetical protein DEHA0C01155g [Debaryomyces hansenii CBS767] 26 Q9Y7B1 60391213 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 27 Q6CKK7 59799028 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 28 NP_011310 6321233 Fatty-acyl coenzyme A oxidase, involved in the fatty acid beta- oxidation pathway; localized to the peroxisomal matrix; Pox1p [Saccharomyces cerevisiae] 29 AAA34891 172217 acyl-coenzyme A oxidase 30 Q6FY63 59799032 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 31 Q756A9 60391210 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 32 CAA68660 2673 acyl-coenzyme A oxidase [Candida tropicalis] 33 1W07B 58177067 Chain B, Arabidopsis Thaliana Acyl-Coa Oxidase 1 34 O65202 62286589 Acyl-coenzyme A oxidase 1, peroxisomal (AOX 1) (Long-chain acyl-CoA oxidase) (AtCX1) 35 NP_001 . . . 55741614 acyl-Coenzyme A oxidase 1, palmitoyl [Gallus gallus] 36 AAH85743 55249685 Acyl-Coenzyme A oxidase 1, palmitoyl [Rattus norvegicus] 37 XP_414406 50754485 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Gallus gallus] 38 H71434 7488059 probable apetala2 domain TINY-Arabidopsis thaliana 39 2DDHA 93279231 Chain A, Crystal Structure Of Acyl-Coa Oxidase Complexed With 3-Oh- Dodecanoate 40 NP_567513 18414744 ACX1 (ACYL-COA OXIDASE 1) [Arabidopsis thaliana] 41 NP_001 . . . 62751552 MGC108278 protein [Xenopus tropicalis] 42 AAI08648 83318302 Unknown (protein for MGC: 131363) [Xenopus laevis] 43 AAH97101 67678187 Zgc: 92584 protein [Danio rerio] 44 AAH63727 39795632 MGC68531 protein [Xenopus laevis] 45 NP_001 . . . 54400372 hypothetical protein LOC449662 [Danio rerio] 46 AAL01888 15553480 acyl-CoA oxidase [Glycine max] 47 ABE83706 92875906 acyl-CoA oxidase [Medicago truncatula] 48 AAL01887 15553478 acyl-CoA oxidase [Glycine max] 49 BAE25794 74188249 unnamed protein product [Mus musculus] 50 NP_056544 66793429 acyl-Coenzyme A oxidase 1, palmitoyl [Mus musculus] 51 XP_001 . . . 109118312 PREDICTED: similar to acyl-Coenzyme A oxidase isoform b [Macaca mulatta] 52 BAA86870 6429156 peroxisomal acyl-CoA oxidase [Mus musculus] 53 Q9R0H0 51338830 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 54 AAB62926 2253380 peroxisomal acyl-CoA oxidase [Mus musculus] 55 AAA18595 495475 peroxisomal fatty acyl-coA oxidase 56 Q5RC19 62286600 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 57 2FONC 109157678 Chain C, X-Ray Crystal Structure Of Leacx1, An Acyl-Coa Oxidase From Lycopersicon Esculentum (Tomato) 58 AAW78689 58531948 peroxisomal acyl-CoA oxidase 1A [Lycopersicon esculentum] 59 AAW78691 58531952 peroxisomal acyl-CoA oxidase 1A [Lycopersicon cheesmaniae] 60 CAH90691 55727877 hypothetical protein [Pongo pygmaeus] 61 NP_009223 30089974 acyl-Coenzyme A oxidase isoform b [Homo sapiens] 62 Q15067 17380467 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) (Straight-chain acyl-CoA oxidase) (SCOX) 63 AAH21339 18204156 Acox2 protein [Mus musculus] 64 NP_004026 30089972 acyl-Coenzyme A oxidase isoform a [Homo sapiens] 65 NP_001 . . . 78369480 acyl-Coenzyme A oxidase 1, palmitoyl [Bos taurus] 66 Q9ZQP2 62286640 Putative acyl-coenzyme A oxidase 1.2, peroxisomal 67 AAH08767 14250616 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 68 AAH68891 46250227 MGC83074 protein [Xenopus laevis] 69 Q8HYL8 34098564 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 70 XP_540441 73965013 PREDICTED: similar to acyl-Coenzyme A oxidase isoform a [Canis familiaris] 71 AAB30019 7689914 peroxisomal acyl-coenzyme A oxidase [Homo sapiens] 72 CAD97622 31873262 hypothetical protein [Homo sapiens] 73 I38095 2117541 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal-human 74 BAF18456 113594582 Os06g0103500 [Oryza sativa (japonica cultivar-group)] 75 AAH10425 14714578 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 76 AAA19113 458119 acyl-CoA oxidase 77 B54942 1082171 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal splice form II- human 78 Q9QXD1 17367045 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 79 Q9Z1N0 17367267 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 80 CAJ83302 89271321 acyl-Coenzyme A oxidase 2, branched chain [Xenopus tropicalis] 81 XP_511690 55645995 PREDICTED: similar to hypothetical protein [Pan troglodytes] 82 XP_592892 76648807 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 1 [Bos taurus] 83 XP_879996 76648809 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 6 [Bos taurus] 84 AAO15577 27462768 acyl-CoA oxidase type 2 [Phascolarctos cinereus] 85 NP_665713 21955130 acyl-Coenzyme A oxidase 2, branched chain [Rattus norvegicus] 86 XP_320718 58394417 ENSANGP00000020032 [Anopheles gambiae str. PEST] 87 NP_510603 17568313 F59F4.1 [Caenorhabditis elegans] 88 XP_541826 73985284 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Canis familiaris] 89 CAE69737 39596101 Hypothetical protein CBG16008 [Caenorhabditis briggsae] 90 CAE63479 39592402 Hypothetical protein CBG07946 [Caenorhabditis briggsae] 91 XP_782824 72015136 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 92 AAH47700 28839704 Acyl-Coenzyme A oxidase 2, branched chain [Homo sapiens] 93 XP_785210 72015040 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 94 EAT39640 108875415 acyl-CoA oxidase [Aedes aegypti] 95 EAS31618 90301987 hypothetical protein CIMG_07097 [Coccidioides immitis RS] 96 O02767 17366131 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 97 NP_998312 47085909 pristanoyl acyl-Coenzyme A oxidase 3 [Danio rerio] 98 XP_783450 72111190 PREDICTED: similar to acyl-Coenzyme A oxidase 1, palmitoyl [Strongylocentrotus purpuratus] 99 EAR95568 89297580 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 100 XP_320717 58394415 ENSANGP00000020118 [Anopheles gambiae str. PEST] 101 NP_493263 17506533 F08A8.3 [Caenorhabditis elegans] 102 AAF73843 8163758 acyl-CoA oxidase ACX3 [Arabidopsis thaliana] 103 BAC26167 26324826 unnamed protein product [Mus musculus] 104 Q9LLH9 62286634 Acyl-coenzyme A oxidase 3, peroxisomal precursor (AOX 3) (Medium-chain acyl-CoA oxidase) (AtCX3) 105 NP_493264 17506535 F08A8.4 [Caenorhabditis elegans] 106 XP_758355 71010163 hypothetical protein UM02208.1 [Ustilago maydis 521] 107 XP_664356 67541164 hypothetical protein AN6752.2 [Aspergillus nidulans FGSC A4] 108 EAT48205 108883980 acyl-CoA oxidase [Aedes aegypti] 109 XP_879827 76648805 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 5 [Bos taurus] 110 NP_499119 17552648 C48B4.1 [Caenorhabditis elegans] 111 O15254 17366151 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 112 BAE55537 83765394 unnamed protein product [Aspergillus oryzae] 113 XP_625136 66556360 PREDICTED: similar to CG5009-PA, partial [Apis mellifera] 114 CAE63476 39592399 Hypothetical protein CBG07943 [Caenorhabditis briggsae] 115 NP_523802 17647123 acyl-Coenzyme A oxidase at 57D proximal CG9707-PA [Drosophila melanogaster] 116 AAH54727 68262425 Acox1 protein [Mus musculus] 117 BAC35782 74197284 unnamed protein product [Mus musculus] 118 AAW78690 58531950 peroxisomal acyl-CoA oxidase 1B [Lycopersicon esculentum] 119 AAL39944 17862934 SD03592p [Drosophila melanogaster] 120 XP_786081 72005647 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 121 ZP_012 . . . 90587482 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Flavobacterium johnsoniae UW102] 122 CAE63477 39592400 Hypothetical protein CBG07944 [Caenorhabditis briggsae] 123 EAR89261 89291273 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 124 CAE65171 39590798 Hypothetical protein CBG10043 [Caenorhabditis briggsae] 125 CAH91120 55728762 hypothetical protein [Pongo pygmaeus] 126 XP_587157 76620513 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Bos taurus] 127 XP_382463 46110811 hypothetical protein FG02287.1 [Gibberella zeae PH-2] 128 XP_973660 91082769 PREDICTED: similar to CG5009-PA [Tribolium castaneum] 129 CAH91864 55730283 hypothetical protein [Pongo pygmaeus] 130 NP_493262 17506531 F08A8.2 [Caenorhabditis elegans] 131 NP_001 . . . 71983346 F08A8.1a [Caenorhabditis elegans] 132 NP_109646 34328334 acyl-Coenzyme A oxidase 3, pristanoyl [Mus musculus] 133 AAH44725 28703869 Acox3 protein [Mus musculus] 134 XP_969513 91093755 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Tribolium castaneum] 135 CAE63480 39592403 Hypothetical protein CBG07947 [Caenorhabditis briggsae] 136 CAJ58493 85539740 Hypothetical protein F08A8.1c [Caenorhabditis elegans] 137 NP_001 . . . 71983355 F08A8.1b [Caenorhabditis elegans] 138 XP_635946 66804335 hypothetical protein DDBDRAFT_0188674 [Dictyostelium discoideum AX4] 139 BAF19505 113595631 Os06g0354500 [Oryza sativa (japonica cultivar-group)] 140 XP_786025 72005645 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 141 NP_611264 24654595 CG5009-PA [Drosophila melanogaster] 142 NP_445791 16758056 acyl-Coenzyme A oxidase 3, pristanoyl [Rattus norvegicus] 143 Q9EPL9 17366740 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 144 Q9LMI7 62286635 Putative acyl-coenzyme A oxidase 3.2, peroxisomal precursor 145 BAE42553 74143079 unnamed protein product [Mus musculus] 146 AAL28144 16767852 GH01266p [Drosophila melanogaster] 147 EAT87650 111066530 hypothetical protein SNOG_05259 [Phaeosphaeria nodorum SN15] 148 BAC26136 26324764 unnamed protein product [Mus musculus] 149 XP_545908 73951751 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Canis familiaris] 150 EAR99631 89301643 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 151 AAC13497 3044212 acyl-CoA oxidase [Arabidopsis thaliana] 152 O65201 62286588 Acyl-coenzyme A oxidase 2, peroxisomal precursor (AOX 2) (Long-chain acyl-CoA oxidase) (AtCX2) 153 AAF14635 6503198 acyl-CoA oxidase [Petroselinum crispum] 154 EAS02716 89304728 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 155 AAR00586 37699750 acyl-CoA oxidase [Phalaenopsis cv. ‘True Lady’] 156 EAS04704 89306716 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 157 YP_634556 108758121 putative acyl-CoA dehydrogenase [Myxococcus xanthus DK 1622] 158 O64894 62286587 Acyl-coenzyme A oxidase, peroxisomal precursor (AOX) (Long-chain acyl-CoA oxidase) 159 AAB67883 1575556 acyl-CoA oxidase homolog [Phalaenopsis sp. ‘True Lady’] 160 ZP_012 . . . 89890388 acyl-CoA oxidase [Flavobacteria bacterium BBFL7] 161 EAL26359 54636956 GA18591-PA [Drosophila pseudoobscura] 162 EAL26647 54637244 GA21980-PA [Drosophila pseudoobscura] 163 EAR98581 89300593 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 164 AAH17053 16877606 ACOX3 protein [Homo sapiens] 165 XP_636605 66805767 hypothetical protein DDBDRAFT_0188084 [Dictyostelium discoideum AX4] 166 YP_445820 83816544 acyl-coenzyme A oxidase I, putative [Salinibacter ruber DSM 13855] 167 EAT87454 111066334 hypothetical protein SNOG_05063 [Phaeosphaeria nodorum SN15] 168 NP_001 . . . 79332306 ACX2 (ACYL-COA OXIDASE 2); acyl-CoA oxidase [Arabidopsis thaliana] 169 YP_290295 72162638 hypothetical protein Tfu_2239 [Thermobifida fusca YX] 170 XP_646395 66826081 hypothetical protein DDBDRAFT_0216722 [Dictyostelium discoideum AX4] 171 XP_317513 58390136 ENSANGP00000010039 [Anopheles gambiae str. PEST] 172 ZP_012 . . . 90205075 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium vanbaalenii PYR-1] 173 EAS36523 90306892 hypothetical protein CIMG_01877 [Coccidioides immitis RS] 174 XP_758113 71007469 hypothetical protein UM01966.1 [Ustilago maydis 521] 175 XP_395486 66564429 PREDICTED: similar to CG9527-PA [Apis mellifera] 176 ABA94670 108864573 Acyl-coenzyme A oxidase 2, peroxisomal precursor, putative, expressed [Oryza sativa (japonica cultivar-group)] 177 XP_637610 66807775 hypothetical protein DDBDRAFT_0187085 [Dictyostelium discoideum AX4] 178 ZP_003 . . . 62426247 COG1960: Acyl-CoA dehydrogenases [Brevibacterium linens BL2] 179 YP_702012 111019040 acyl-CoA oxidase [Rhodococcus sp. RHA1] 180 NP_500943 17540842 F58F9.7 [Caenorhabditis elegans] 181 EAT39346 108875121 acyl-CoA oxidase [Aedes aegypti] 182 CAA96917 1945301 POX1 [Saccharomyces cerevisiae] 183 NP_609027 45550141 CG9527-PA [Drosophila melanogaster] 184 CAA04688 2370232 putative acyl-CoA oxidase [Hordeum vulgare subsp. vulgare] 185 AAL48010 17944231 LD22081p [Drosophila melanogaster] 186 EAL32329 54643586 GA18278-PA [Drosophila pseudoobscura] 187 CAE61577 39584202 Hypothetical protein CBG05491 [Caenorhabditis briggsae] 188 AAL48950 17945806 RE34879p [Drosophila melanogaster] 189 NP_724181 24585195 CG17544-PC, isoform C [Drosophila melanogaster] 190 AAL14003 16186117 SD05719p [Drosophila melanogaster] 191 XP_317446 58390030 ENSANGP00000011863 [Anopheles gambiae str. PEST] 192 ZP_011 . . . 89339305 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium flavescens PYR-GCK] 193 ZP_004 . . . 66967405 Acyl-CoA oxidase: Acyl-CoA dehydrogenase, C-terminal [Arthrobacter sp. FB24] 194 ZP_011 . . . 88803915 acyl-coenzyme A oxidase I, putative [Robiginitalea biformata HTCC2501] 195 EAL32912 54644170 GA21857-PA [Drosophila pseudoobscura] 196 EAL26648 54637245 GA21981-PA [Drosophila pseudoobscura] 197 ZP_012 . . . 92908524 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium sp. JLS]

TABLE 38 Examples of pox3 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_503244 50551539 YlPOX3 [Yarrowia lipolytica] 2 XP_502199 50549457 YlPOX5 [Yarrowia lipolytica] 3 O74935 59799073 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) 4 O74934 59799072 Acyl-coenzyme A oxidase 1 (Acyl-CoA oxidase 1) 5 XP_504703 50554589 YlPOX1 [Yarrowia lipolytica] 6 XP_504475 50554133 YlPOX4 [Yarrowia lipolytica] 7 XP_503632 50552444 hypothetical protein [Yarrowia lipolytica] 8 XP_571768 58269224 Acyl-coenzyme A oxidase I [Cryptococcus neoformans var. neoformans JEC21] 9 XP_760471 71020481 hypothetical protein UM04324.1 [Ustilago maydis 521] 10 XP_717995 68475844 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 11 Q6BRD5 59799027 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 12 BAA83482 5763520 acyl-CoA oxidase [Candida tropicalis] 13 OXCKPM 66060 acyl-CoA oxidase (EC 1.3.3.6) PXP4, peroxisomal-yeast (Candida maltosa) 14 BAA83483 5763522 acyl-CoA oxidase [Candida tropicalis] 15 AAA34362 170912 acyl-coenzyme A oxidase II precursor 16 P08790 17373073 Acyl-coenzyme A oxidase 5 (Acyl-CoA oxidase 5) (PXP-5) 17 OXCKX5 66061 acyl-CoA oxidase (EC 1.3.3.6) POX5, peroxisomal-yeast (Candida tropicalis) 18 XP_721610 68468582 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 19 Q9Y7B1 60391213 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 20 XP_457726 50417804 hypothetical protein DEHA0C01155g [Debaryomyces hansenii CBS767] 21 P06598 17373023 Acyl-coenzyme A oxidase 4 (Acyl-CoA oxidase 4) (PXP-4) (Peroxisomal fatty acyl-CoA oxidase) 22 AAA34361 170910 PXP-2 protein 23 1306283A 225549 oxidase, fatty acyl 24 XP_721613 68468588 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 25 XP_459232 50421365 hypothetical protein DEHA0D18667g [Debaryomyces hansenii CBS767] 26 Q00468 2498206 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) (AOX 2) 27 Q6FY63 59799032 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 28 Q6CKK7 59799028 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 29 Q756A9 60391210 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 30 NP_011310 6321233 Fatty-acyl coenzyme A oxidase, involved in the fatty acid beta- oxidation pathway; localized to the peroxisomal matrix; Pox1p [Saccharomyces cerevisiae] 31 AAA34891 172217 acyl-coenzyme A oxidase 32 CAA68660 2673 acyl-coenzyme A oxidase [Candida tropicalis] 33 BAF18456 113594582 Os06g0103500 [Oryza sativa (japonica cultivar-group)] 34 O65202 62286589 Acyl-coenzyme A oxidase 1, peroxisomal (AOX 1) (Long-chain acyl-CoA oxidase) (AtCX1) 35 H71434 7488059 probable apetala2 domain TINY-Arabidopsis thaliana 36 1W07B 58177067 Chain B, Arabidopsis Thaliana Acyl-Coa Oxidase 1 37 NP_567513 18414744 ACX1 (ACYL-COA OXIDASE 1) [Arabidopsis thaliana] 38 AAH68891 46250227 MGC83074 protein [Xenopus laevis] 39 AAL01888 15553480 acyl-CoA oxidase [Glycine max] 40 ABE83706 92875906 acyl-CoA oxidase [Medicago truncatula] 41 Q9ZQP2 62286640 Putative acyl-coenzyme A oxidase 1.2, peroxisomal 42 CAJ83302 89271321 acyl-Coenzyme A oxidase 2, branched chain [Xenopus tropicalis] 43 BAE25794 74188249 unnamed protein product [Mus musculus] 44 AAL01887 15553478 acyl-CoA oxidase [Glycine max] 45 AAW78689 58531948 peroxisomal acyl-CoA oxidase 1A [Lycopersicon esculentum] 46 2FONC 109157678 Chain C, X-Ray Crystal Structure Of Leacx1, An Acyl-Coa Oxidase From Lycopersicon Esculentum (Tomato) 47 AAW78691 58531952 peroxisomal acyl-CoA oxidase 1A [Lycopersicon cheesmaniae] 48 AAH21339 18204156 Acox2 protein [Mus musculus] 49 XP_320717 58394415 ENSANGP00000020118 [Anopheles gambiae str. PEST] 50 Q9QXD1 17367045 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 51 XP_414406 50754485 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Gallus gallus] 52 NP_001 . . . 54400372 hypothetical protein LOC449662 [Danio rerio] 53 AAH63727 39795632 MGC68531 protein [Xenopus laevis] 54 EAT39640 108875415 acyl-CoA oxidase [Aedes aegypti] 55 AAH97101 67678187 Zgc: 92584 protein [Danio rerio] 56 NP_001 . . . 55741614 acyl-Coenzyme A oxidase 1, palmitoyl [Gallus gallus] 57 NP_001 . . . 62751552 MGC108278 protein [Xenopus tropicalis] 58 NP_665713 21955130 acyl-Coenzyme A oxidase 2, branched chain [Rattus norvegicus] 59 XP_320718 58394417 ENSANGP00000020032 [Anopheles gambiae str. PEST] 60 AAI08648 83318302 Unknown (protein for MGC: 131363) [Xenopus laevis] 61 CAE63479 39592402 Hypothetical protein CBG07946 [Caenorhabditis briggsae] 62 Q8HYL8 34098564 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 63 EAS31618 90301987 hypothetical protein CIMG_07097 [Coccidioides immitis RS] 64 CAH90691 55727877 hypothetical protein [Pongo pygmaeus] 65 Q5RC19 62286600 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 66 XP_001 . . . 109118312 PREDICTED: similar to acyl-Coenzyme A oxidase isoform b [Macaca mulatta] 67 XP_540441 73965013 PREDICTED: similar to acyl-Coenzyme A oxidase isoform a [Canis familiaris] 68 I38095 2117541 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal-human 69 AAH08767 14250616 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 70 NP_004026 30089972 acyl-Coenzyme A oxidase isoform a [Homo sapiens] 71 CAD97622 31873262 hypothetical protein [Homo sapiens] 72 NP_510603 17568313 F59F4.1 [Caenorhabditis elegans] 73 Q15067 17380467 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) (Straight-chain acyl-CoA oxidase) (SCOX) 74 NP_009223 30089974 acyl-Coenzyme A oxidase isoform b [Homo sapiens] 75 NP_001 . . . 78369480 acyl-Coenzyme A oxidase 1, palmitoyl [Bos taurus] 76 B54942 1082171 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal splice form II- human 77 AAA18595 495475 peroxisomal fatty acyl-coA oxidase 78 AAA19113 458119 acyl-CoA oxidase 79 AAB30019 7689914 peroxisomal acyl-coenzyme A oxidase [Homo sapiens] 80 XP_758355 71010163 hypothetical protein UM02208.1 [Ustilago maydis 521] 81 AAH10425 14714578 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 82 CAE65171 39590798 Hypothetical protein CBG10043 [Caenorhabditis briggsae] 83 O02767 17366131 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 84 AAH47700 28839704 Acyl-Coenzyme A oxidase 2, branched chain [Homo sapiens] 85 AAO15577 27462768 acyl-CoA oxidase type 2 [Phascolarctos cinereus] 86 CAE69737 39596101 Hypothetical protein CBG16008 [Caenorhabditis briggsae] 87 2DDHA 93279231 Chain A, Crystal Structure Of Acyl-Coa Oxidase Complexed With 3-Oh- Dodecanoate 88 NP_499119 17552648 C48B4.1 [Caenorhabditis elegans] 89 XP_625136 66556360 PREDICTED: similar to CG5009-PA, partial [Apis mellifera] 90 XP_879996 76648809 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 6 [Bos taurus] 91 XP_592892 76648807 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 1 [Bos taurus] 92 XP_782824 72015136 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 93 Q9LLH9 62286634 Acyl-coenzyme A oxidase 3, peroxisomal precursor (AOX 3) (Medium-chain acyl-CoA oxidase) (AtCX3) 94 AAF73843 8163758 acyl-CoA oxidase ACX3 [Arabidopsis thaliana] 95 AAH85743 55249685 Acyl-Coenzyme A oxidase 1, palmitoyl [Rattus norvegicus] 96 XP_785210 72015040 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 97 XP_511690 55645995 PREDICTED: similar to hypothetical protein [Pan troglodytes] 98 NP_493263 17506533 F08A8.3 [Caenorhabditis elegans] 99 XP_973660 91082769 PREDICTED: similar to CG5009-PA [Tribolium castaneum] 100 CAE63477 39592400 Hypothetical protein CBG07944 [Caenorhabditis briggsae] 101 NP_056544 66793429 acyl-Coenzyme A oxidase 1, palmitoyl [Mus musculus] 102 Q9Z1N0 17367267 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 103 EAT48205 108883980 acyl-CoA oxidase [Aedes aegypti] 104 BAA86870 6429156 peroxisomal acyl-CoA oxidase [Mus musculus] 105 Q9R0H0 51338830 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 106 EAR95568 89297580 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 107 AAB62926 2253380 peroxisomal acyl-CoA oxidase [Mus musculus] 108 AAW78690 58531950 peroxisomal acyl-CoA oxidase 1B [Lycopersicon esculentum] 109 XP_635946 66804335 hypothetical protein DDBDRAFT_0188674 [Dictyostelium discoideum AX4] 110 NP_493264 17506535 F08A8.4 [Caenorhabditis elegans] 111 CAE63480 39592403 Hypothetical protein CBG07947 [Caenorhabditis briggsae] 112 NP_001 . . . 71983346 F08A8.1a [Caenorhabditis elegans] 113 CAE63476 39592399 Hypothetical protein CBG07943 [Caenorhabditis briggsae] 114 BAC26167 26324826 unnamed protein product [Mus musculus] 115 XP_541826 73985284 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Canis familiaris] 116 BAE55537 83765394 unnamed protein product [Aspergillus oryzae] 117 NP_001 . . . 71983355 F08A8.1b [Caenorhabditis elegans] 118 CAJ58493 85539740 Hypothetical protein F08A8.1c [Caenorhabditis elegans] 119 EAS04704 89306716 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 120 NP_493262 17506531 F08A8.2 [Caenorhabditis elegans] 121 EAT87650 111066530 hypothetical protein SNOG_05259 [Phaeosphaeria nodorum SN15] 122 O64894 62286587 Acyl-coenzyme A oxidase, peroxisomal precursor (AOX) (Long-chain acyl-CoA oxidase) 123 XP_664356 67541164 hypothetical protein AN6752.2 [Aspergillus nidulans FGSC A4] 124 XP_783450 72111190 PREDICTED: similar to acyl-Coenzyme A oxidase 1, palmitoyl [Strongylocentrotus purpuratus] 125 BAF19505 113595631 Os06g0354500 [Oryza sativa (japonica cultivar-group)] 126 XP_382463 46110811 hypothetical protein FG02287.1 [Gibberella zeae PH-1] 127 XP_879827 76648805 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 5 [Bos taurus] 128 Q9LMI7 62286635 Putative acyl-coenzyme A oxidase 3.2, peroxisomal precursor 129 AAR00586 37699750 acyl-CoA oxidase [Phalaenopsis cv. ‘True Lady’] 130 CAH91864 55730283 hypothetical protein [Pongo pygmaeus] 131 NP_998312 47085909 pristanoyl acyl-Coenzyme A oxidase 3 [Danio rerio] 132 AAL39944 17862934 SD03592p [Drosophila melanogaster] 133 NP_523802 17647123 acyl-Coenzyme A oxidase at 57D proximal CG9707-PA [Drosophila melanogaster] 134 EAR99631 89301643 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 135 AAB67883 1575556 acyl-CoA oxidase homolog [Phalaenopsis sp. ‘True Lady’] 136 AAL48010 17944231 LD22081p [Drosophila melanogaster] 137 EAR98581 89300593 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 138 YP_445820 83816544 acyl-coenzyme A oxidase I, putative [Salinibacter ruber DSM 13855] 139 EAL26648 54637245 GA21981-PA [Drosophila pseudoobscura] 140 O65201 62286588 Acyl-coenzyme A oxidase 2, peroxisomal precursor (AOX 2) (Long-chain acyl-CoA oxidase) (AtCX2) 141 AAC13497 3044212 acyl-CoA oxidase [Arabidopsis thaliana] 142 AAF14635 6503198 acyl-CoA oxidase [Petroselinum crispum] 143 XP_636605 66805767 hypothetical protein DDBDRAFT_0188084 [Dictyostelium discoideum AX4] 144 O15254 17366151 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 145 XP_758113 71007469 hypothetical protein UM01966.1 [Ustilago maydis 521] 146 NP_611264 24654595 CG5009-PA [Drosophila melanogaster] 147 XP_969513 91093755 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Tribolium castaneum] 148 NP_109646 34328334 acyl-Coenzyme A oxidase 3, pristanoyl [Mus musculus] 149 XP_637610 66807775 hypothetical protein DDBDRAFT_0187085 [Dictyostelium discoideum AX4] 150 BAC35782 74197284 unnamed protein product [Mus musculus] 151 EAL26647 54637244 GA21980-PA [Drosophila pseudoobscura] 152 AAH44725 28703869 Acox3 protein [Mus musculus] 153 AAH54727 68262425 Acox1 protein [Mus musculus] 154 XP_587157 76620513 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Bos taurus] 155 ZP_012 . . . 90587482 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Flavobacterium johnsoniae UW101] 156 CAA04688 2370232 putative acyl-CoA oxidase [Hordeum vulgare subsp. vulgare] 157 EAL26359 54636956 GA18591-PA [Drosophila pseudoobscura] 158 AAL28144 16767852 GH01266p [Drosophila melanogaster] 159 CAH91120 55728762 hypothetical protein [Pongo pygmaeus] 160 NP_445791 16758056 acyl-Coenzyme A oxidase 3, pristanoyl [Rattus norvegicus] 161 Q9EPL9 17366740 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 162 XP_786081 72005647 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 163 EAR89261 89291273 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 164 ZP_012 . . . 89890388 acyl-CoA oxidase [Flavobacteria bacterium BBFL7] 165 XP_646395 66826081 hypothetical protein DDBDRAFT_0216722 [Dictyostelium discoideum AX4] 166 NP_001 . . . 79332306 ACX2 (ACYL-COA OXIDASE 2); acyl-CoA oxidase [Arabidopsis thaliana] 167 BAC26136 26324764 unnamed protein product [Mus musculus] 168 BAE42553 74143079 unnamed protein product [Mus musculus] 169 EAS02716 89304728 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 170 ABA94670 108864573 Acyl-coenzyme A oxidase 2, peroxisomal precursor, putative, expressed [Oryza sativa (japonica cultivar-group)] 171 YP_290295 72162638 hypothetical protein Tfu_2239 [Thermobifida fusca YX] 172 EAL32329 54643586 GA18278-PA [Drosophila pseudoobscura] 173 EAT87454 111066334 hypothetical protein SNOG_05063 [Phaeosphaeria nodorum SN15] 174 YP_634556 1087581 putative acyl-CoA dehydrogenase [Myxococcus xanthus DK 211622] 175 NP_508036 17560134 F25C8.1 [Caenorhabditis elegans] 176 NP_572371 24640268 CG4586-PA [Drosophila melanogaster] 177 ZP_012 . . . 90205075 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium vanbaalenii PYR-2] 178 XP_545908 73951751 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Canis familiaris] 179 YP_702012 111019040 acyl-CoA oxidase [Rhodococcus sp. RHA1] 180 ZP_011 . . . 89339305 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium flavescens PYR-GCK] 181 XP_786025 72005645 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 182 BAE60166 83770031 unnamed protein product [Aspergillus oryzae] 183 ZP_012 . . . 92908524 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium sp. JLS] 184 NP_961035 41408199 hypothetical protein MAP2101 [Mycobacterium avium subsp. paratuberculosis K-10] 185 YP_640598 108800401 acyl-CoA dehydrogenase-like protein [Mycobacterium sp. MCS] 186 XP_317446 58390030 ENSANGP00000011863 [Anopheles gambiae str. PEST] 187 EAT48670 108884445 acyl-CoA oxidase [Aedes aegypti] 188 BAE47462 78483008 acyl-CoA oxidase [Arthrobacter ureafaciens] 189 ZP_003 . . . 62426247 COG1960: Acyl-CoA dehydrogenases [Brevibacterium linens BL2] 190 AAH17053 16877606 ACOX3 protein [Homo sapiens] 191 ZP_011 . . . 88854811 acyl-CoA oxidase [marine actinobacterium PHSC20C2] 192 ZP_006 . . . 71369913 Acyl-CoA oxidase: Acyl-CoA dehydrogenase, C-terminal: Acyl- CoA dehydrogenase, central region: Acyl-CoA dehydrogenase, N-terminal [Nocardioides sp. J5614] 193 ZP_009 . . . 84496319 hypothetical protein JNB_02330 [Janibacter sp. HTCC2649] 194 ZP_004 . . . 66967405 Acyl-CoA oxidase: Acyl-CoA dehydrogenase, C-terminal [Arthrobacter sp. FB24] 195 ZP_011 . . . 88803915 acyl-coenzyme A oxidase I, putative [Robiginitalea biformata HTCC2501] 196 CAA68661 2674 44 kD translation product [Candida tropicalis]

TABLE 39 Examples of pox5 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_502199 50549457 YlPOX5 [Yarrowia lipolytica]. 2 O74936 59799074 Acyl-coenzyme A oxidase 3 (Acyl-CoA oxidase 3) 3 O74934 59799072 Acyl-coenzyme A oxidase 1 (Acyl-CoA oxidase 1) 4 XP_504703 50554589 YlPOX1 [Yarrowia lipolytica] 5 O74935 59799073 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) 6 XP_504475 50554133 YlP0X4 [Yarrowia lipolytica] 7 XP_503632 50552444 hypothetical protein [Yarrowia lipolytica] 8 XP_760471 71020481 hypothetical protein UM04324.1 [Ustilago maydis 521] 9 XP_571768 58269224 Acyl-coenzyme A oxidase I [Cryptococcus neoformans var. neoformans JEC21] 10 BAA83482 5763520 acyl-CoA oxidase [Candida tropicalis] 11 BAA83483 5763522 acyl-CoA oxidase [Candida tropicalis] 12 OXCKPM 66060 acyl-CoA oxidase (EC 1.3.3.6) PXP4, peroxisomal-yeast (Candida maltosa) 13 P08790 17373073 Acyl-coenzyme A oxidase 5 (Acyl-CoA oxidase 5) (PXP-5) 14 AAA34362 170912 acyl-coenzyme A oxidase II precursor 15 OXCKX5 66061 acyl-CoA oxidase (EC 1.3.3.6) POX5, peroxisomal-yeast (Candida tropicalis) 16 XP_717995 68475844 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 17 Q9Y7B1 60391213 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 18 XP_721610 68468582 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 19 AAA34361 170910 PXP-2 protein 20 XP_721613 68468588 putative fatty-acyl coenzyme A oxidase [Candida albicans SC5314] 21 P06598 17373023 Acyl-coenzyme A oxidase 4 (Acyl-CoA oxidase 4) (PXP-4) (Peroxisomal fatty acyl-CoA oxidase) 22 Q6BRD5 59799027 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 23 1306283A 225549 oxidase, fatty acyl 24 XP_457726 50417804 hypothetical protein DEHA0C01155g [Debaryomyces hansenii CBS767] 25 XP_459232 50421365 hypothetical protein DEHA0D18667g [Debaryomyces hansenii CBS767] 26 Q00468 2498206 Acyl-coenzyme A oxidase 2 (Acyl-CoA oxidase 2) (AOX 2) 27 Q756A9 60391210 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 28 Q6FY63 59799032 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 29 NP_011310 6321233 Fatty-acyl coenzyme A oxidase, involved in the fatty acid beta- oxidation pathway; localized to the peroxisomal matrix; Pox1p [Saccharomyces cerevisiae] 30 AAA34891 172217 acyl-coenzyme A oxidase 31 Q6CKK7 59799028 Acyl-coenzyme A oxidase (Acyl-CoA oxidase) 32 1W07B 58177067 Chain B, Arabidopsis Thaliana Acyl-Coa Oxidase 1 33 O65202 62286589 Acyl-coenzyme A oxidase 1, peroxisomal (AOX 1) (Long-chain acyl-CoA oxidase) (AtCX1) 34 CAA68660 2673 acyl-coenzyme A oxidase [Candida tropicalis] 35 NP_567513 18414744 ACX1 (ACYL-COA OXIDASE 1) [Arabidopsis thaliana] 36 H71434 7488059 probable apetala2 domain TINY-Arabidopsis thaliana 37 AAL01888 15553480 acyl-CoA oxidase [Glycine max] 38 AAL01887 15553478 acyl-CoA oxidase [Glycine max] 39 ABE83706 92875906 acyl-CoA oxidase [Medicago truncatula] 40 BAF18456 113594582 Os06g0103500 [Oryza sativa (japonica cultivar-group)] 41 2FONC 109157678 Chain C, X-Ray Crystal Structure Of Leacx1, An Acyl-Coa Oxidase From Lycopersicon Esculentum (Tomato) 42 AAW78689 58531948 peroxisomal acyl-CoA oxidase 1A [Lycopersicon esculentum] 43 AAW78691 58531952 peroxisomal acyl-CoA oxidase 1A [Lycopersicon cheesmaniae] 44 Q9ZQP2 62286640 Putative acyl-coenzyme A oxidase 1.2, peroxisomal 45 NP_001 . . . 54400372 hypothetical protein LOC449662 [Danio rerio] 46 XP_592892 76648807 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 1 [Bos taurus] 47 XP_879996 76648809 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 6 [Bos taurus] 48 AAH68891 46250227 MGC83074 protein [Xenopus laevis] 49 AAH97101 67678187 Zgc: 92584 protein [Danio rerio] 50 XP_414406 50754485 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Gallus gallus] 51 NP_001 . . . 55741614 acyl-Coenzyme A oxidase 1, palmitoyl [Gallus gallus] 52 XP_320717 58394415 ENSANGP00000020118 [Anopheles gambiae str. PEST] 53 NP_510603 17568313 F59F4.1 [Caenorhabditis elegans] 54 BAE25794 74188249 unnamed protein product [Mus musculus] 55 CAE69737 39596101 Hypothetical protein CBG16008 [Caenorhabditis briggsae] 56 AAH21339 18204156 Acox2 protein [Mus musculus] 57 CAJ83302 89271321 acyl-Coenzyme A oxidase 2, branched chain [Xenopus tropicalis] 58 XP_320718 58394417 ENSANGP00000020032 [Anopheles gambiae str. PEST] 59 Q9QXD1 17367045 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 60 AAH63727 39795632 MGC68531 protein [Xenopus laevis] 61 Q5RC19 62286600 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 62 AAW78690 58531950 peroxisomal acyl-CoA oxidase 1B [Lycopersicon esculentum] 63 XP_782824 72015136 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 64 AAH47700 28839704 Acyl-Coenzyme A oxidase 2, branched chain [Homo sapiens] 65 EAT39640 108875415 acyl-CoA oxidase [Aedes aegypti] 66 NP_001 . . . 62751552 MGC108278 protein [Xenopus tropicalis] 67 CAH90691 55727877 hypothetical protein [Pongo pygmaeus] 68 XP_001 . . . 109118312 PREDICTED: similar to acyl-Coenzyme A oxidase isoform b [Macaca mulatta] 69 Q15067 17380467 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) (Straight-chain acyl-CoA oxidase) (SCOX) 70 NP_009223 30089974 acyl-Coenzyme A oxidase isoform b [Homo sapiens] 71 AAA19113 458119 acyl-CoA oxidase 72 AAB30019 7689914 peroxisomal acyl-coenzyme A oxidase [Homo sapiens] 73 XP_785210 72015040 PREDICTED: similar to Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) [Strongylocentrotus purpuratus] 74 O02767 17366131 Acyl-coenzyme A oxidase 2, peroxisomal (3-alpha,7-alpha,12- alpha-trihydroxy-5-beta-cholestanoyl-CoA 24-hydroxylase) (3- alpha,7-alpha,12-alpha-trihydroxy-5-beta-cholestanoyl-CoA oxidase) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA-CoA oxidase) 75 NP_004026 30089972 acyl-Coenzyme A oxidase isoform a [Homo sapiens] 76 AAA18595 495475 peroxisomal fatty acyl-coA oxidase 77 I38095 2117541 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal-human 78 AAH08767 14250616 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 79 B54942 1082171 acyl-CoA oxidase (EC 1.3.3.6), peroxisomal splice form II- human 80 NP_001 . . . 78369480 acyl-Coenzyme A oxidase 1, palmitoyl [Bos taurus] 81 NP_499119 17552648 C48B4.1 [Caenorhabditis elegans] 82 NP_665713 21955130 acyl-Coenzyme A oxidase 2, branched chain [Rattus norvegicus] 83 CAD97622 31873262 hypothetical protein [Homo sapiens] 84 BAA86870 6429156 peroxisomal acyl-CoA oxidase [Mus musculus] 85 NP_056544 66793429 acyl-Coenzyme A oxidase 1, palmitoyl [Mus musculus] 86 AAH85743 55249685 Acyl-Coenzyme A oxidase 1, palmitoyl [Rattus norvegicus] 87 2DDHA 93279231 Chain A, Crystal Structure Of Acyl-Coa Oxidase Complexed With 3-Oh- Dodecanoate 88 Q9R0H0 51338830 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 89 XP_758355 71010163 hypothetical protein UM02208.1 [Ustilago maydis 521] 90 AAH10425 14714578 Acyl-Coenzyme A oxidase 1, palmitoyl [Homo sapiens] 91 AAB62926 2253380 peroxisomal acyl-CoA oxidase [Mus musculus] 92 CAE65171 39590798 Hypothetical protein CBG10043 [Caenorhabditis briggsae] 93 XP_625136 66556360 PREDICTED: similar to CG5009-PA, partial [Apis mellifera] 94 XP_540441 73965013 PREDICTED: similar to acyl-Coenzyme A oxidase isoform a [Canis familiaris] 95 Q8HYL8 34098564 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 96 CAE63476 39592399 Hypothetical protein CBG07943 [Caenorhabditis briggsae] 97 AAI08648 83318302 Unknown (protein for MGC: 131363) [Xenopus laevis] 98 XP_879827 76648805 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) isoform 5 [Bos taurus] 99 AAF73843 8163758 acyl-CoA oxidase ACX3 [Arabidopsis thaliana] 100 Q9LLH9 62286634 Acyl-coenzyme A oxidase 3, peroxisomal precursor (AOX 3) (Medium-chain acyl-CoA oxidase) (AtCX3) 101 XP_973660 91082769 PREDICTED: similar to CG5009-PA [Tribolium castaneum] 102 EAS31618 90301987 hypothetical protein C1MG_07097 [Coccidioides immitis RS] 103 EAR95568 89297580 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 104 Q9Z1N0 17367267 Acyl-coenzyme A oxidase 1, peroxisomal (Palmitoyl-CoA oxidase) (AOX) 105 AAO15577 27462768 acyl-CoA oxidase type 2 [Phascolarctos cinereus] 106 XP_541826 73985284 PREDICTED: similar to Acyl-coenzyme A oxidase 2, peroxisomal (Branched-chain acyl-CoA oxidase) (BRCACox) (Trihydroxycoprostanoyl-CoA oxidase) (THCCox) (THCA- CoA oxidase) [Canis familiaris] 107 CAE63479 39592402 Hypothetical protein CBG07946 [Caenorhabditis briggsae] 108 XP_511690 55645995 PREDICTED: similar to hypothetical protein [Pan troglodytes] 109 BAC26167 26324826 unnamed protein product [Mus musculus] 110 CAE63477 39592400 Hypothetical protein CBG07944 [Caenorhabditis briggsae] 111 NP_493264 17506535 F08A8.4 [Caenorhabditis elegans] 112 Q9LMI7 62286635 Putative acyl-coenzyme A oxidase 3.2, peroxisomal precursor 113 CAJ58493 85539740 Hypothetical protein F08A8.1c [Caenorhabditis elegans] 114 NP_001 . . . 71983355 F08A8.1b [Caenorhabditis elegans] 115 NP_001 . . . 71983346 F08A8.1a [Caenorhabditis elegans] 116 BAF19505 113595631 Os06g0354500 [Oryza sativa (japonica cultivar-group)] 117 NP_493263 17506533 F08A8.3 [Caenorhabditis elegans] 118 O64894 62286587 Acyl-coenzyme A oxidase, peroxisomal precursor (AOX) (Long-chain acyl-CoA oxidase) 119 NP_493262 17506531 F08A8.2 [Caenorhabditis elegans] 120 CAE63480 39592403 Hypothetical protein CBG07947 [Caenorhabditis briggsae] 121 AAF14635 6503198 acyl-CoA oxidase [Petroselinum crispum] 122 BAE55537 83765394 unnamed protein product [Aspergillus oryzae] 123 NP_998312 47085909 pristanoyl acyl-Coenzyme A oxidase 3 [Danio rerio] 124 O65201 62286588 Acyl-coenzyme A oxidase 2, peroxisomal precursor (AOX 2) (Long-chain acyl-CoA oxidase) (AtCX2) 125 AAC13497 3044212 acyl-CoA oxidase [Arabidopsis thaliana] 126 EAT87650 111066530 hypothetical protein SNOG_05259 [Phaeosphaeria nodorum SN15] 127 XP_587157 76620513 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Bos taurus] 128 EAT48205 108883980 acyl-CoA oxidase [Aedes aegypti] 129 EAT87454 111066334 hypothetical protein SNOG_05063 [Phaeosphaeria nodorum SN15] 130 XP_664356 67541164 hypothetical protein AN6752.2 [Aspergillus nidulans FGSC A4] 131 AAB67883 1575556 acyl-CoA oxidase homolog [Phalaenopsis sp. ‘True Lady’] 132 AAR00586 37699750 acyl-CoA oxidase [Phalaenopsis cv. ‘True Lady’] 133 CAH91864 55730283 hypothetical protein [Pongo pygmaeus] 134 XP_636605 66805767 hypothetical protein DDBDRAFT_0188084 [Dictyostelium discoideum AX4] 135 NP_611264 24654595 CG5009-PA [Drosophila melanogaster] 136 EAS04704 89306716 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 137 XP_635946 66804335 hypothetical protein DDBDRAFT_0188674 [Dictyostelium discoideum AX4] 138 O15254 17366151 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 139 AAL39944 17862934 SD03592p [Drosophila melanogaster] 140 NP_523802 17647123 acyl-Coenzyme A oxidase at 57D proximal CG9707-PA [Drosophila melanogaster] 141 AAL28144 16767852 GH01266p [Drosophila melanogaster] 142 XP_382463 46110811 hypothetical protein FG02287.1 [Gibberella zeae PH-2] 143 EAL26359 54636956 GA18591-PA [Drosophila pseudoobscura] 144 CAA04688 2370232 putative acyl-CoA oxidase [Hordeum vulgare subsp. vulgare] 145 XP_783450 72111190 PREDICTED: similar to acyl-Coenzyme A oxidase 1, palmitoyl [Strongylocentrotus purpuratus] 146 AAL48010 17944231 LD22081p [Drosophila melanogaster] 147 CAH91120 55728762 hypothetical protein [Pongo pygmaeus] 148 XP_646395 66826081 hypothetical protein DDBDRAFT_0216722 [Dictyostelium discoideum AX4] 149 XP_758113 71007469 hypothetical protein UM01966.1 [Ustilago maydis 521] 150 EAR99631 89301643 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 151 EAR89261 89291273 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 152 XP_637610 66807775 hypothetical protein DDBDRAFT_0187085 [Dictyostelium discoideum AX4] 153 ABA94670 108864573 Acyl-coenzyme A oxidase 2, peroxisomal precursor, putative, expressed [Oryza sativa (japonica cultivar-group)] 154 EAL26647 54637244 GA21980-PA [Drosophila pseudoobscura] 155 ZP_012 . . . 90587482 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Flavobacterium johnsoniae UW102] 156 BAC35782 74197284 unnamed protein product [Mus musculus] 157 EAS02716 89304728 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 158 YP_634556 1087581 putative acyl-CoA dehydrogenase [Myxococcus xanthus DK 211622] 159 AAH54727 68262425 Acox1 protein [Mus musculus] 160 YP_445820 83816544 acyl-coenzyme A oxidase I, putative [Salinibacter ruber DSM 13855] 161 AAH44725 28703869 Acox3 protein [Mus musculus] 162 NP_109646 34328334 acyl-Coenzyme A oxidase 3, pristanoyl [Mus musculus] 163 XP_545908 73951751 PREDICTED: similar to Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) [Canis familiaris] 164 NP_001 . . . 79332306 ACX2 (ACYL-COA OXIDASE 2); acyl-CoA oxidase [Arabidopsis thaliana] 165 EAR98581 89300593 Acyl-CoA oxidase family protein [Tetrahymena thermophila SB210] 166 XP_969513 91093755 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Tribolium castaneum] 167 ZP_012 . . . 89890388 acyl-CoA oxidase [Flavobacteria bacterium BBFL7] 168 YP_290295 72162638 hypothetical protein Tfu_2239 [Thermobifida fusca YX] 169 BAC26136 26324764 unnamed protein product [Mus musculus] 170 BAE42553 74143079 unnamed protein product [Mus musculus] 171 EAL26648 54637245 GA21981-PA [Drosophila pseudoobscura] 172 NP_445791 16758056 acyl-Coenzyme A oxidase 3, pristanoyl [Rattus norvegicus] 173 Q9EPL9 17366740 Acyl-coenzyme A oxidase 3, peroxisomal (Pristanoyl-CoA oxidase) (Branched-chain acyl-CoA oxidase) (BRCACox) 174 XP_786025 72005645 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 175 XP_786081 72005647 PREDICTED: similar to pristanoyl acyl-Coenzyme A oxidase 3 [Strongylocentrotus purpuratus] 176 EAL32329 54643586 GA18278-PA [Drosophila pseudoobscura] 177 NP_724181 24585195 CG17544-PC, isoform C [Drosophila melanogaster] 178 AAL14003 16186117 SD05719p [Drosophila melanogaster] 179 CAA68661 2674 44 kD translation product [Candida tropicalis] 180 NP_572371 24640268 CG4586-PA [Drosophila melanogaster] 181 ZP_011 . . . 88854811 acyl-CoA oxidase [marine actinobacterium PHSC20C1] 182 EAL33335 54644594 GA14550-PA [Drosophila pseudoobscura] 183 EAS36523 90306892 hypothetical protein CIMG_01877 [Coccidioides immitis RS] 184 EAT48670 108884445 acyl-CoA oxidase [Aedes aegypti] 185 YP_702012 111019040 acyl-CoA oxidase [Rhodococcus sp. RHA1] 186 ZP_003 . . . 62426247 COG1960: Acyl-CoA dehydrogenases [Brevibacterium linens BL2] 187 XP_317446 58390030 ENSANGP00000011863 [Anopheles gambiae str. PEST] 188 XP_317513 58390136 ENSANGP00000010039 [Anopheles gambiae str. PEST] 189 AAH17053 16877606 ACOX3 protein [Homo sapiens] 190 ZP_012 . . . 90205075 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium vanbaalenii PYR-1] 191 ZP_011 . . . 88803915 acyl-coenzyme A oxidase I, putative [Robiginitalea biformata HTCC2502] 192 XP_642669 66817952 hypothetical protein DDBDRAFT_0169270 [Dictyostelium discoideum AX4] 193 NP_508036 17560134 F25C8.1 [Caenorhabditis elegans] 194 ZP_006 . . . 71369913 Acyl-CoA oxidase: Acyl-CoA dehydrogenase, C-terminal: Acyl- CoA dehydrogenase, central region: Acyl-CoA dehydrogenase, N-terminal [Nocardioides sp. JS614] 195 BAE65934 83775815 unnamed protein product [Aspergillus oryzae] 196 ZP_012 . . . 92908524 Acyl-CoA oxidase-like: Acyl-CoA dehydrogenase-like: Acyl- CoA dehydrogenase, central region [Mycobacterium sp. JLS]

TABLE 40 Examples of triglyceride lipase (tgl4) polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_505230 50555644 hypothetical protein [Yarrowia lipolytica]. 2 XP_459553 50421985 hypothetical protein DEHA0E06127g [Debaryomyces hansenii CBS767] 3 XP_387731 46126355 hypothetical protein FG07555.1 [Gibberella zeae PH-1] 4 XP_717704 68476486 putative patatin-like phospholipase [Candida albicans SC5314] 5 XP_958750 85091127 hypothetical protein [Neurospora crassa OR74A] 6 NP_013015 6322942 Triacylglycerol lipase involved in triacylglycerol mobilization and degradation; found in lipid particles; potential Cdc28p substrate; Tgl4p [Saccharomyces cerevisiae] 7 XP_659317 67522513 hypothetical protein AN1713.2 [Aspergillus nidulans FGSC A4] 8 NP_984327 45188104 ADR231Cp [Eremothecium gossypii] 9 CAD60564 27764284 unnamed protein product [Podospora anserina] 10 EAT92134 111071014 hypothetical protein SNOG_00639 [Phaeosphaeria nodorum SN15] 11 BAE59228 83769091 unnamed protein product [Aspergillus oryzae] 12 XP_446465 50288073 unnamed protein product [Candida glabrata] 13 XP_751982 70994310 patatin-like serine hydrolase [Aspergillus fumigatus Af293] 14 XP_451336 50302799 unnamed protein product [Kluyveromyces lactis] 15 EAS30095 90300464 hypothetical protein CIMG_08841 [Coccidioides immitis RS] 16 EAQ83092 88175624 hypothetical protein CHGG_10910 [Chaetomium globosum CBS 148.51] 17 NP_014724 6324655 Triacylglycerol lipase involved in TAG mobilization; localizes to lipid particles; potential Cdc28p substrate; Tgl5p [Saccharomyces cerevisiae] 18 XP_448470 50292075 unnamed protein product [Candida glabrata] 19 ABG79933 110628937 triacylglycerol lipase [Magnaporthe grisea] 20 NP_593197 19114109 hypothetical protein SPAC1A6.05c [Schizosaccharomyces pombe 972h-] 21 EAR86065 89288077 Patatin-like phospholipase family protein [Tetrahymena thermophila SB210] 22 XP_463448 50902030 P0512C01.22 [Oryza sativa (japonica cultivar-group)] 23 BAF06239 113533856 Os01g0762000 [Oryza sativa (japonica cultivar-group)] 24 YP_433865 83645430 predicted esterase of the alpha-beta hydrolase superfamily [Hahella chejuensis KCTC 2396] 25 ZP_008 . . . 77951643 conserved hypothetical protein [Marinobacter aquaeolei VT8] 26 AAM97135 22531263 putative protein [Arabidopsis thaliana] 27 BAF13581 113550138 Os03g0810900 [Oryza sativa (japonica cultivar-group)] 28 ABE78088 92867978 phospholipase, patatin family, putative [Medicago truncatula] 29 YP_693301 110834442 hypothetical protein ABO_1581 [Alcanivorax borkumensis SK2] 30 NP_974449 42572707 unknown protein [Arabidopsis thaliana] 31 BAE56507 83766364 unnamed protein product [Aspergillus oryzae] 32 EAR83738 89285729 Patatin-like phospholipase family protein [Tetrahymena thermophila SB210] 33 ABE94194 92898449 Patatin [Medicago truncatula] 34 XP_752278 70995024 patatin-like serine hydrolase [Aspergillus fumigatus Af293] 35 YP_046026 50084516 hypothetical protein ACIAD1335 [Acinetobacter sp. ADP1] 36 ZP_011 . . . 88794068 predicted esterase of the alpha-beta hydrolase superfamily protein [Alteromonas macleodii ‘Deep ecotype’] 37 XP_380696 46107274 hypothetical protein FG00520.1 [Gibberella zeae PH-1] 38 ZP_011 . . . 88703608 Patatin-like phospholipase [gamma proteobacterium KT 71] 39 EAT85718 111064598 hypothetical protein SNOG_07067 [Phaeosphaeria nodorum SN15] 40 ZP_011 . . . 88707125 Patatin-like phospholipase [gamma proteobacterium KT 71] 41 XP_757164 71004996 hypothetical protein UM01017.1 [Ustilago maydis 521] 42 YP_615253 103485692 Patatin [Sphingopyxis alaskensis RB2256] 43 XP_964502 85113293 hypothetical protein [Neurospora crassa OR74A] 44 NP_594022 63054585 hypothetical protein SPAC31G5.20c [Schizosaccharomyces pombe 972h-] 45 XP_502898 50550851 hypothetical protein [Yarrowia lipolytica] 46 XP_722550 68466709 putative patatin-like phospholipase [Candida albicans SC5314] 47 EAS31687 90302056 hypothetical protein CIMG_07166 [Coccidioides immitis RS] 48 BAE57969 83767830 unnamed protein product [Aspergillus oryzae] 49 XP_722412 68466994 putative patatin-like phospholipase [Candida albicans SC5314] 50 EAQ93362 88185894 hypothetical protein CHGG_01597 [Chaetomium globosum CBS 148.51] 51 ZP_010 . . . 85708568 hypothetical protein NAP1_04995 [Erythrobacter sp. NAP1] 52 XP_457141 50412575 hypothetical protein DEHA0B04048g [Debaryomyces hansenii CBS767] 53 XP_658012 67516253 hypothetical protein AN0408.2 [Aspergillus nidulans FGSC A4] 54 EAS33873 90304242 hypothetical protein CIMG_04897 [Coccidioides immitis RS] 55 XP_805126 71404931 hypothetical protein [Trypanosoma cruzi strain CL Brener] 56 CAB88226 7630152 conserved hypothetical protein L5213T.05 [Leishmania major] 57 CAJ06433 68127889 hypothetical protein, conserved [Leishmania major] 58 XP_750246 70990794 patatin-like serine hydrolase [Aspergillus fumigatus Af293] 59 ABG79934 110628939 triacylglycerol lipase [Magnaporthe grisea] 60 XP_386821 46124535 hypothetical protein FG06645.1 [Gibberella zeae PH-1] 61 EAT92413 111071293 hypothetical protein SNOG_00918 [Phaeosphaeria nodorum SN15] 62 NP_588315 19075815 hypothetical protein SPCC1450.16c [Schizosaccharomyces pombe 972h-] 63 EAQ90965 88183497 hypothetical protein CHGG_02900 [Chaetomium globosum CBS 148.51] 64 XP_957267 85084210 hypothetical protein [Neurospora crassa OR74A] 65 XP_451445 50303015 unnamed protein product [Kluyveromyces lactis] 66 CAB11704 2388971 SPAC31G5.20c [Schizosaccharomyces pombe] 67 XP_626587 66358818 hypothetical protein cgd2_4050 [Cryptosporidium parvum Iowa II] 68 XP_665308 67588018 hypothetical protein Chro.20432 [Cryptosporidium hominis TU502] 69 XP_449952 50295082 hypothetical protein CAGL0M13981g [Candida glabrata CBS138] 70 XP_722764 68466564 putative lipid particle triacylglycerol lipase fragment [Candida albicans SC5314] 71 NP_986330 45200760 AGL337Cp [Eremothecium gossypii] 72 CAE84405 39722369 Tgl3 protein [Kluyveromyces delphensis] 73 XP_807283 71409928 hypothetical protein [Trypanosoma cruzi strain CL Brener] 74 XP_807016 71409330 hypothetical protein [Trypanosoma cruzi strain CL Brener] 75 XP_502944 50550941 hypothetical protein [Yarrowia lipolytica] 76 NP_014044 6323973 Triacylglycerol lipase of the lipid particle, responsible for all the TAG lipase activity of the lipid particle; contains the consensus sequence motif GXSXG, which is found in lipolytic enzymes; Tgl3p [Saccharomyces cerevisiae]

TABLE 41 Examples of triglyceride lipase (tgl3) polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_502944 50550941 hypothetical protein [Yarrowia lipolytica]. 2 XP_752278 70995024 patatin-like serine hydrolase [Aspergillus fumigatus Af293] 3 BAE56507 83766364 unnamed protein product [Aspergillus oryzae] 4 EAT85718 111064598 hypothetical protein SNOG_07067 [Phaeosphaeria nodorum SN15] 5 XP_380696 46107274 hypothetical protein FG00520.1 [Gibberella zeae PH-1] 6 XP_964502 85113293 hypothetical protein [Neurospora crassa OR74A] 7 EAQ93362 88185894 hypothetical protein CHGG_01597 [Chaetomium globosum CBS 148.51] 8 XP_722764 68466564 putative lipid particle triacylglycerol lipase fragment [Candida albicans SC5314] 9 NP_986330 45200760 AGL337Cp [Eremothecium gossypii] 10 CAE84405 39722369 Tgl3 protein [Kluyveromyces delphensis] 11 XP_449952 50295082 hypothetical protein CAGL0M13981g [Candida glabrata CBS138] 12 XP_451445 50303015 unnamed protein product [Kluyveromyces lactis] 13 NP_014044 6323973 Triacylglycerol lipase of the lipid particle, responsible for all the TAG lipase activity of the lipid particle; contains the consensus sequence motif GXSXG, which is found in lipolytic enzymes; Tgl3p [Saccharomyces cerevisiae] 14 EAS31687 90302056 hypothetical protein CIMG_07166 [Coccidioides immitis RS] 15 EAT92134 111071014 hypothetical protein SNOG_00639 [Phaeosphaeria nodorum SN15] 16 XP_387731 46126355 hypothetical protein FG07555.1 [Gibberella zeae PH-1] 17 NP_588315 19075815 hypothetical protein SPCC1450.16c [Schizosaccharomyces pombe 972h-] 18 XP_751982 70994310 patatin-like serine hydrolase [Aspergillus fumigatus Af293] 19 XP_958750 85091127 hypothetical protein [Neurospora crassa OR74A] 20 EAQ83092 88175624 hypothetical protein CHGG_10910 [Chaetomium globosum CBS 148.51] 21 BAE59228 83769091 unnamed protein product [Aspergillus oryzae] 22 YP_693301 110834442 hypothetical protein ABO_1581 [Alcanivorax borkumensis SK2] 23 XP_659317 67522513 hypothetical protein AN1713.2 [Aspergillus nidulans FGSC A4] 24 ZP_011 . . . 88703608 Patatin-like phospholipase [gamma proteobacterium KT 71] 25 BAF06239 113533856 Os01g0762000 [Oryza sativa (japonica cultivar-group)] 26 XP_463448 50902030 P0512C01.22 [Oryza sativa (japonica cultivar-group)] 27 EAS30095 90300464 hypothetical protein CIMG_08841 [Coccidioides immitis RS] 28 YP_433865 83645430 predicted esterase of the alpha-beta hydrolase superfamily [Hahella chejuensis KCTC 2396] 29 XP_459553 50421985 hypothetical protein DEHA0E06127g [Debaryomyces hansenii CBS767] 30 CAD60564 27764284 unnamed protein product [Podospora anserina] 31 ZP_008 . . . 77951643 conserved hypothetical protein [Marinobacter aquaeolei VT8] 32 XP_505230 50555644 hypothetical protein [Yarrowia lipolytica] 33 XP_446465 50288073 unnamed protein product [Candida glabrata] 34 AAM97135 22531263 putative protein [Arabidopsis thaliana] 35 BAF13581 113550138 Os03g0810900 [Oryza sativa (japonica cultivar-group)] 36 ABG79933 110628937 triacylglycerol lipase [Magnaporthe grisea] 37 XP_717704 68476486 putative patatin-like phospholipase [Candida albicans SC5314] 38 NP_013015 6322942 Triacylglycerol lipase involved in triacylglycerol mobilization and degradation; found in lipid particles; potential Cdc28p substrate; Tgl4p [Saccharomyces cerevisiae] 39 NP_594022 63054585 hypothetical protein SPAC31G5.20c [Schizosaccharomyces pombe 972h-] 40 EAR83738 89285729 Patatin-like phospholipase family protein [Tetrahymena thermophila SB210] 41 NP_984327 45188104 ADR231Cp [Eremothecium gossypii] 42 EAR86065 89288077 Patatin-like phospholipase family protein [Tetrahymena thermophila SB210] 43 NP_014724 6324655 Triacylglycerol lipase involved in TAG mobilization; localizes to lipid particles; potential Cdc28p substrate; Tgl5p [Saccharomyces cerevisiae] 44 YP_046026 50084516 hypothetical protein ACIAD1335 [Acinetobacter sp. ADP1] 45 XP_451336 50302799 unnamed protein product [Kluyveromyces lactis] 46 ZP_011 . . . 88794068 predicted esterase of the alpha-beta hydrolase superfamily protein [Alteromonas macleodii ‘Deep ecotype’] 47 XP_722550 68466709 putative patatin-like phospholipase [Candida albicans SC5314] 48 XP_722412 68466994 putative patatin-like phospholipase [Candida albicans SC5314] 49 EAS33873 90304242 hypothetical protein CIMG_04897 [Coccidioides immitis RS] 50 ABG79934 110628939 triacylglycerol lipase [Magnaporthe grisea] 51 ZP_011 . . . 88707125 Patatin-like phospholipase [gamma proteobacterium KT 71] 52 NP_593197 19114109 hypothetical protein SPAC1A6.05c [Schizosaccharomyces pombe 972h-] 53 YP_615253 103485692 Patatin [Sphingopyxis alaskensis RB2256] 54 ZP_010 . . . 85708568 hypothetical protein NAP1_04995 [Erythrobacter sp. NAP1] 55 XP_658012 67516253 hypothetical protein AN0408.2 [Aspergillus nidulans FGSC A4] 56 XP_448470 50292075 unnamed protein product [Candida glabrata] 57 XP_807283 71409928 hypothetical protein [Trypanosoma cruzi strain CL Brener] 58 XP_386821 46124535 hypothetical protein FG06645.1 [Gibberella zeae PH-1]

TABLE 42 Examples of acyl-coA-cholesterol polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_505086.1 50555355 hypothetical protein [Yarrowia lipolytica]. 2 XP_449806 50294790 hypothetical protein CAGL0M10571g [Candida glabrata CBS138] 3 AAO32554 28564940 ARE2 [Saccharomyces kluyveri] 4 XP_461395 50425599 hypothetical protein DEHA0F25652g [Debaryomyces hansenii CBS767] 5 P84285 56404462 Sterol O-acyltransferase 2 (Sterol-ester synthase) (ASAT) 6 XP_714776 68482533 acyl-CoA cholesterol acyltransferase [Candida albicans SC5314] 7 AAO32474 28564191 ARE2 [Saccharomyces castellii] 8 NP_982606 45184888 AAR065Cp [Eremothecium gossypii] 9 NP_014416 6324346 Acyl-CoA:sterol acyltransferase, isozyme of Are1p; endoplasmic reticulum enzyme that contributes the major sterol esterification activity in the presence of oxygen; Are2p [Saccharomyces cerevisiae] 10 AAC49441 1389739 acyl-CoA:sterol acyltransferase 11 XP_452607 50305295 unnamed protein product [Kluyveromyces lactis] 12 Q876L2 34582301 Sterol O-acyltransferase 2 (Sterol-ester synthase 2) 13 Q876L3 34582302 Sterol O-acyltransferase 1 (Sterol-ester synthase 1) 14 AAT92940 51013293 YCR048W [Saccharomyces cerevisiae] 15 XP_445307 50285757 unnamed protein product [Candida glabrata] 16 NP_593707 19114619 hypothetical protein SPAC13G7.05 [Schizosaccharomyces pombe 972h-] 17 XP_370268 39977761 hypothetical protein MG06765.4 [Magnaporthe grisea 70-15] 18 EAQ84619 88177151 hypothetical protein CHGG_08633 [Chaetomium globosum CBS 148.51] 19 XP_961300 85102248 related to acyl-CoA sterol acyltransferase [MIPS] [Neurospora crassa OR74A] 20 XP_750354 70991010 sterol o-acyltransferase APE2 [Aspergillus fumigatus Af293] 21 EAT80792 111059672 hypothetical protein SNOG_11748 [Phaeosphaeria nodorum SN15] 22 XP_661812 67527926 hypothetical protein AN4208.2 [Aspergillus nidulans FGSC A4] 23 BAE54934 83764790 unnamed protein product [Aspergillus oryzae] 24 EAT89521 111068401 predicted protein [Phaeosphaeria nodorum SN15] 25 EAQ86094 88178626 hypothetical protein CHGG_07347 [Chaetomium globosum CBS 148.51] 26 NP_588558 19076058 hypothetical protein SPCP1E11.05c [Schizosaccharomyces pombe 972h-] 27 XP_956576 85080625 hypothetical protein [Neurospora crassa OR74A] 28 EAS32815 90303184 hypothetical protein CIMG_03839 [Coccidioides immitis RS] 29 XP_382192 46110268 hypothetical protein FG02016.1 [Gibberella zeae PH-1] 30 XP_761502 71022545 hypothetical protein UM05355.1 [Ustilago maydis 521] 31 EAL20032 50257323 hypothetical protein CNBF3580 [Cryptococcus neoformans var. neoformans B-3501A] 32 XP_571260 58268208 sterol O-acyltransferase [Cryptococcus neoformans var. neoformans JEC21] 33 EAR83561 89285549 MBOAT family protein [Tetrahymena thermophila SB210] 34 EAS03424 89305436 MBOAT family protein [Tetrahymena thermophila SB210] 35 AAS72375 45642961 acyl-CoA:cholesterol acyltransferase alpha [Toxoplasma gondii] 36 AAS72376 45642963 acyl-CoA:cholesterol acyltransferase beta [Toxoplasma gondii] 37 XP_640280 111226542 hypothetical protein DDBDRAFT_0205259 [Dictyostelium discoideum AX4] 38 XP_645633 66824557 hypothetical protein DDBDRAFT_0202877 [Dictyostelium discoideum AX4] 39 EAS35506 90305875 hypothetical protein CIMG_00860 [Coccidioides immitis RS] 40 XP_386864 46124621 hypothetical protein FG06688.1 [Gibberella zeae PH-1] 41 NP_649816 28571583 CG8112-PA [Drosophila melanogaster] 42 CAE75170 39591950 Hypothetical protein CBG23107 [Caenorhabditis briggsae] 43 NP_033256 84619697 sterol O-acyltransferase 1 [Mus musculus] 44 BAC34925 26342537 unnamed protein product [Mus musculus] 45 XP_359451 39939828 hypothetical protein MG05326.4 [Magnaporthe grisea 70-15] 46 Q61263 18202591 Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (Acyl coenzyme A:cholesterol acyltransferase 1) (ACAT-1) 47 XP_975142 91083363 PREDICTED: similar to CG31991-PA, isoform A [Tribolium castaneum] 48 EAL28962 54639560 GA20833-PA [Drosophila pseudoobscura] 49 XP_957022 85082953 hypothetical protein [Neurospora crassa OR74A] 50 XP_547445 73961286 PREDICTED: similar to Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (Acyl coenzyme A:cholesterol acyltransferase 1) (ACAT-1) [Canis familiaris] 51 EAQ89124 88181656 hypothetical protein CHGG_05743 [Chaetomium globosum CBS 148.51] 52 EAT78735 111057615 hypothetical protein SNOG_13711 [Phaeosphaeria nodorum SN15] 53 Q60457 18202585 Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (Acyl coenzyme A:cholesterol acyltransferase 1) (ACAT-1) 54 XP_624754 110762262 PREDICTED: similar to midway CG31991-PA, isoform A [Apis mellifera] 55 O70536 18202126 Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (Acyl coenzyme A:cholesterol acyltransferase 1) (ACAT-1) 56 XP_728984 82915156 hypothetical protein PY01256 [Plasmodium yoelii yoelii str. 17XNL] 57 NP_505828 71997360 H19N07.4 [Caenorhabditis elegans] 58 NP_001 . . . 77735363 hypothetical protein LOC504287 [Bos taurus] 59 2201440A 1585676 acyl-CoA/cholesterol acyltransferase 60 XP_663763 67539978 hypothetical protein AN6159.2 [Aspergillus nidulans FGSC A4] 61 CAI13574 55960156 sterol O-acyltransferase (acyl-Coenzyme A: cholesterol acyltransferase) 1 [Homo sapiens] 62 XP_514030 55588858 PREDICTED: hypothetical protein XP_514030 [Pan troglodytes] 63 AAL56227 18028942 cholesterol acyltransferase 1 [Gorilla gorilla] 64 AAL56228 18028944 cholesterol acyltransferase 1 [Pongo pygmaeus] 65 AAC37532 4878022 acyl-coenzyme A: cholesterol acyltransferase [Homo sapiens] 66 EAT47694 108883469 sterol o-acyltransferase [Aedes aegypti] 67 XP_390039 46136695 hypothetical protein FG09863.1 [Gibberella zeae PH-1] 68 BAE01048 67969393 unnamed protein product [Macaca fascicularis] 69 O77761 18202178 Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (Acyl coenzyme A:cholesterol acyltransferase 1) (ACAT-1) 70 AAH06263 34782946 DGAT1 protein [Homo sapiens] 71 XP_317656 31226099 ENSANGP00000002281 [Anopheles gambiae str. PEST] 72 XP_320320 58393809 ENSANGP00000016486 [Anopheles gambiae str. PEST] 73 XP_320321 58393811 ENSANGP00000016512 [Anopheles gambiae str. PEST] 74 BAE65302 83775179 unnamed protein product [Aspergillus oryzae] 75 NP_724017 24584734 midway CG31991-PC, isoform C [Drosophila melanogaster] 76 NP_995724 45552403 midway CG31991-PD, isoform D [Drosophila melanogaster] 77 EAL33593 54644853 GA16599-PA [Drosophila pseudoobscura] 78 NP_586145 19074639 STEROL O-ACYLTRANSFERASE [Encephalitozoon cuniculi GB-M1] 79 XP_502557 50550169 hypothetical protein [Yarrowia lipolytica] 80 XP_885914 76617939 PREDICTED: similar to Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (Acyl coenzyme A:cholesterol acyltransferase 2) (ACAT-2) isoform 2 [Bos taurus] 81 XP_591251 76617937 PREDICTED: similar to Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (Acyl coenzyme A:cholesterol acyltransferase 2) (ACAT-2) isoform 1 [Bos taurus] 82 XP_624691 66564061 PREDICTED: similar to CG8112-PA [Apis mellifera] 83 O75908 18202149 Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (Acyl coenzyme A:cholesterol acyltransferase 2) (ACAT-2) 84 XP_001 . . . 109096888 PREDICTED: sterol O-acyltransferase 2 [Macaca mulatta] 85 AAK48829 13898623 acyl coenzyme A: cholesterol acyltransferase-2 [Homo sapiens] 86 AAH77916 50416229 Soat1-prov protein [Xenopus laevis] 87 AAH96091 64654094 Sterol O-acyltransferase 2 [Homo sapiens] 88 EAT38531 108874306 sterol o-acyltransferase [Aedes aegypti] 89 AAH96090 64652990 Sterol O-acyltransferase 2 [Homo sapiens] 90 NP_001 . . . 50539976 hypothetical protein LOC436731 [Danio rerio] 91 NP_714950 40254723 sterol O-acyltransferase 2 [Rattus norvegicus] 92 XP_422267 50751122 PREDICTED: similar to Sterol O-acyltransferase 1 (Cholesterol acyltransferase 1) (Acyl coenzyme A:cholesterol acyltransferase 1) (ACAT-1) [Gallus gallus] 93 O88908 18202245 Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (Acyl coenzyme A:cholesterol acyltransferase 2) (ACAT-2) 94 O77759 18202176 Sterol O-acyltransferase 2 (Cholesterol acyltransferase 2) (Acyl coenzyme A:cholesterol acyltransferase 2) (ACAT-2) 95 NP_666176 22122547 sterol O-acyltransferase 2 [Mus musculus] 96 BAC00846 21392392 AcylCoA:Cholesterol Acyltransferase 2 [Rattus norvegicus] 97 XP_692855 68364838 PREDICTED: similar to Soat1-prov protein [Danio rerio] 98 XP_543637 73996435 PREDICTED: similar to sterol O-acyltransferase 2 [Canis familiaris] 99 CAG11944 47225461 unnamed protein product [Tetraodon nigroviridis] 100 CAF96514 47222847 unnamed protein product [Tetraodon nigroviridis] 101 CAG10815 47219451 unnamed protein product [Tetraodon nigroviridis] 102 EAR94647 89296659 MBOAT family protein [Tetrahymena thermophila SB210] 103 AAP68322 31711932 At2g19450 [Arabidopsis thaliana] 104 EAR94441 89296453 MBOAT family protein [Tetrahymena thermophila SB210] 105 CAG07990 47225647 unnamed protein product [Tetraodon nigroviridis] 106 XP_788209 72085563 PREDICTED: similar to sterol O-acyltransferase 1 [Strongylocentrotus purpuratus] 107 AAO32475 28564193 ARE2 [Saccharomyces castellii] 108 XP_673128 68062248 hypothetical protein PB300300.00.0 [Plasmodium berghei strain ANKA]

TABLE 43 Examples of phospholipid-diacylglycerol polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_504038 50553256 hypothetical protein [Yarrowia lipolytica]. 2 XP_458897 50420721 hypothetical protein DEHA0D10868g [Debaryomyces hansenii CBS767] 3 XP_718878 68473963 hypothetical protein CaO19_13439 [Candida albicans SC5314] 4 XP_447501 50290139 hypothetical protein CAGL0I05786g [Candida glabrata CBS138] 5 XP_455369 50310695 unnamed protein product [Kluyveromyces lactis] 6 NP_014405 6324335 Acyltransferase that catalyzes diacylglycerol esterification; one of several acyltransferases that contribute to triglyceride synthesis; putative homolog of human lecithin cholesterol acyltransferase; Lro1p [Saccharomyces cerevisiae] 7 NP_985371 45198342 AFL179Cp [Eremothecium gossypii] 8 BAE57684 83767545 unnamed protein product [Aspergillus oryzae] 9 EAS32928 90303297 hypothetical protein CIMG_03952 [Coccidioides immitis RS] 10 EAT89812 111068692 hypothetical protein SNOG_03081 [Phaeosphaeria nodorum SN15] 11 XP_662596 67537644 hypothetical protein AN4992.2 [Aspergillus nidulans FGSC A4] 12 XP_361017 39942960 hypothetical protein MG03560.4 [Magnaporthe grisea 70-15] 13 CAD60714 27803011 unnamed protein product [Podospora anserina] 14 XP_718980 68473754 hypothetical protein CaO19_6018 [Candida albicans SC5314] 15 XP_386551 46123995 hypothetical protein FG06375.1 [Gibberella zeae PH-1] 16 XP_959598 85092964 hypothetical protein [Neurospora crassa OR74A] 17 XP_754624 70999814 phospholipid:diacylglycerol acyltransferase [Aspergillus fumigatus Af293] 18 O94680 21362753 Phospholipid:diacylglycerol acyltransferase (PDAT) (Pombe LRO1 homolog 1) 19 EAQ92213 88184745 hypothetical protein CHGG_00448 [Chaetomium globosum CBS 148.51] 20 XP_756469 71003606 hypothetical protein UM00322.1 [Ustilago maydis 521] 21 EAL18953 50256226 hypothetical protein CNBI2140 [Cryptococcus neoformans var. neoformans B-3501A] 22 XP_568014 58261208 phospholipid:diacylglycerol acyltransferase [Cryptococcus neoformans var. neoformans JEC21] 23 AAP47635 37785695 lecithine cholesterol acyltransferase-like protein [Medicago truncatula] 24 AAS79590 45935132 putative phosphatidylcholine-sterol acyltransferase [Ipomoea trifida] 25 BAF25212 113631531 Os09g0444200 [Oryza sativa (japonica cultivar-group)] 26 AAK96619 15450695 AT5g13640/MSH12_10 [Arabidopsis thaliana] 27 XP_829379 74025626 phospholipid:diacylglycerol acyltransferase-like protein [Trypanosoma brucei TREU927] 28 XP_816552 71655997 phospholipid:diacylglycerol acyltransferase [Trypanosoma cruzi strain CL Brener] 29 AAO17787 37724553 lecithine cholesterol acyltransferase-like protein [Arabidopsis thaliana] 30 XP_821756 71660075 phospholipid:diacylglycerol acyltransferase [Trypanosoma cruzi strain CL Brener] 31 CAD38153 27552462 putative phosphatidylcholine-sterol acetyltransferase [Physcomitrella patens] 32 NP_190069 15230521 phosphatidylcholine-sterol O-acyltransferase [Arabidopsis thaliana] 33 CAD38155 27552466 putative acetyltransferase [Physcomitrella patens] 34 CAJ02843 68124573 phospholipid:diacylglycerol acyltransferase, putative [Leishmania major]

TABLE 44 Examples of malate dehydrogenase polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_503933.1 50553046 hypothetical protein [Yarrowia lipolytica]. 2 EAT83219 111062099 predicted protein [Phaeosphaeria nodorum SN15] 3 XP_747556 70984070 malate dehydrogenase, NAD-dependent [Aspergillus fumigatus Af293] 4 BAE61776 83771646 unnamed protein product [Aspergillus oryzae] 5 EAQ89010 88181542 malate dehydrogenase, mitochondrial precursor [Chaetomium globosum CBS 148.51] 6 EAS37226 90307595 hypothetical protein CIMG_02580 [Coccidioides immitis RS] 7 XP_572013 58269714 L-malate dehydrogenase [Cryptococcus neoformans var. neoformans JEC21] 8 XP_748936 70986899 malate dehydrogenase, NAD-dependent [Aspergillus fumigatus Af293] 9 BAE55502 83765359 unnamed protein product [Aspergillus oryzae] 10 EAQ88553 88181085 malate dehydrogenase, mitochondrial precursor [Chaetomium globosum CBS 148.51] 11 ABH10641 111606553 malate dehydrogenase [Coccidioides posadasii] 12 XP_321163 58395331 ENSANGP00000020184 [Anopheles gambiae str. PEST] 13 XP_717047 68477790 putative peroxisomal malate dehydrogenase [Candida albicans SC5314] 14 AAY63978 67043759 mitochondrial malate dehydrogenase [Lysiphlebus testaceipes] 15 AAX19496 60393102 mitochondrial malate dehydrogenase 2b [Xenopus laevis] 16 EAT40089 108875864 malate dehydrogenase [Aedes aegypti] 17 AAI06696 76780392 Mdh2a protein [Xenopus laevis] 18 XP_572038 58269764 malate dehydrogenase [Cryptococcus neoformans var. neoformans JEC21] 19 XP_392478 66513092 PREDICTED: similar to mitochondrial malate dehydrogenase precursor isoform 1 [Apis mellifera] 20 XP_973533 91085015 PREDICTED: similar to mitochondrial malate dehydrogenase precursor [Tribolium castaneum] 21 CAG12894 47224065 unnamed protein product [Tetraodon nigroviridis] 22 NP_001 . . . 58332672 mitochondrial malate dehydrogenase 2 [Xenopus tropicalis] 23 XP_819104 71664243 malate dehydrogenase [Trypanosoma cruzi strain CL Brener] 24 CAJ07715 68129178 malate dehydrogenase [Leishmania major] 25 EAL29124 54639722 GA20754-PA [Drosophila pseudoobscura] 26 XP_708782 68367713 PREDICTED: similar to malate dehydrogenase, mitochondrial isoform 6 [Danio rerio] 27 XP_708779 68367700 PREDICTED: similar to malate dehydrogenase, mitochondrial isoform 3 [Danio rerio] 28 AAW79319 58613465 malate dehydrogenase [Isochrysis galbana] 29 CAI29601 56403598 hypothetical protein [Pongo pygmaeus] 30 XP_460754 50424335 hypothetical protein DEHA0F09911g [Debaryomyces hansenii CBS767] 31 XP_001 . . . 109066259 PREDICTED: similar to mitochondrial malate dehydrogenase precursor [Macaca mulatta] 32 AAK69767 14583133 malate dehydrogenase [Sphyraena idiastes] 33 XP_708778 68367696 PREDICTED: similar to malate dehydrogenase, mitochondrial isoform 2 [Danio rerio] 34 NP_001 . . . 79327392 PMDH2 [Arabidopsis thaliana] 35 ABE84799 92877831 Malate dehydrogenase, NAD-dependent, eukaryotes and gamma proteobacteria [Medicago truncatula] 36 AAU29200 52139820 glyoxisomal malate dehydrogenase [Lycopersicon esculentum] 37 AAW27425 56758570 SJCHGC06124 protein [Schistosoma japonicum] 38 XP_708781 68367709 PREDICTED: similar to malate dehydrogenase, mitochondrial isoform 5 [Danio rerio] 39 1SMKH 60593494 Chain H, Mature And Translocatable Forms Of Glyoxysomal Malate Dehydrogenase Have Different Activities And Stabilities But Similar Crystal Structures 40 XP_822509 71746908 mitochondrial malate dehydrogenase [Trypanosoma brucei TREU927] 41 1SEVB 60593476 Chain B, Mature And Translocatable Forms Of Glyoxysomal Malate Dehydrogenase Have Different Activities And Stabilities But Similar Crystal Structures 42 XP_708780 68367705 PREDICTED: similar to malate dehydrogenase, mitochondrial isoform 4 [Danio rerio] 43 ABD77290 89574129 mitochondrial malate dehydrogenase 2, NAD [Homo sapiens] 44 ABD77284 89574117 mitochondrial malate dehydrogenase 2, NAD [Rattus norvegicus] 45 ABD77283 89574115 mitochondrial malate dehydrogenase 2, NAD [Mus musculus] 46 ABD77287 89574123 mitochondrial malate dehydrogenase 2, NAD [Oryctolagus cuniculus] 47 ABD77280 89574109 mitochondrial malate dehydrogenase 2, NAD [Loxodonta africana] 48 ABD77288 89574125 mitochondrial malate dehydrogenase 2, NAD [Lepus europaeus] 49 XP_792004 72025672 PREDICTED: similar to mitochondrial malate dehydrogenase precursor [Strongylocentrotus purpuratus] 50 ABD77293 89574135 mitochondrial malate dehydrogenase 2, NAD [Canis familiaris] 51 ABD77294 89574137 mitochondrial malate dehydrogenase 2, NAD [Felis catus] 52 ABD77278 89574105 mitochondrial malate dehydrogenase 2, NAD [Didelphis virginiana] 53 XP_809210 71414199 mitochondrial malate dehydrogenase [Trypanosoma cruzi strain CL Brener] 54 ABD77301 89574151 mitochondrial malate dehydrogenase 2, NAD [Sus scrofa] 55 ABD77286 89574121 mitochondrial malate dehydrogenase 2, NAD [Mesocricetus auratus] 56 ABD77298 89574145 mitochondrial malate dehydrogenase 2, NAD [Bos taurus] 57 AAT35230 47531133 mitochondrial malate dehydrogenase [Clonorchis sinensis] 58 ABD77281 89574111 mitochondrial malate dehydrogenase 2, NAD [Dasypus novemcinctus] 59 ABD77296 89574141 mitochondrial malate dehydrogenase 2, NAD [Ceratotherium simum] 60 ABA99939 108863014 Malate dehydrogenase, glyoxysomal precursor, putative, expressed [Oryza sativa (japonica cultivar-group)] 61 AAU29198 52139816 mitochondrial malate dehydrogenase [Lycopersicon esculentum] 62 ABD77279 89574107 mitochondrial malate dehydrogenase 2, NAD [Sminthopsis douglasi] 63 CAJ07717 68129180 malate dehydrogenase, putative [Leishmania major] 64 XP_519160 55628772 PREDICTED: similar to mitochondrial malate dehydrogenase precursor [Pan troglodytes] 65 ABD72702 89473780 putative mitochondrial malate dehydrogenase [Acyrthosiphon pisum] 66 ABD77299 89574147 mitochondrial malate dehydrogenase 2, NAD [Balaenoptera physalus] 67 BAF23792 113623847 Os08g0434300 [Oryza sativa (japonica cultivar-group)] 68 CAI11361 57337458 putative malate dehydrogenase [Orpinomyces sp. OUS1] 69 ABD77289 89574127 mitochondrial malate dehydrogenase 2, NAD [Tadarida brasiliensis] 70 BAF06605 113534222 Os01g0829800 [Oryza sativa (japonica cultivar-group)] 71 ABD77292 89574133 mitochondrial malate dehydrogenase 2, NAD [Tupaia glis] 72 ABD77285 89574119 mitochondrial malate dehydrogenase 2, NAD [Cavia porcellus] 73 BAF18303 113579940 Os05g0574400 [Oryza sativa (japonica cultivar-group)] 74 ZP_010 . . . 85712491 malate dehydrogenase [Idiomarina baltica OS145] 75 XP_539718 57097207 PREDICTED: similar to malate dehydrogenase 2, NAD (mitochondrial) [Canis familiaris] 76 ABD77277 89574103 mitochondrial malate dehydrogenase 2, NAD [Monodelphis domestica] 77 Q5R030 62286970 Malate dehydrogenase 78 YP_660122 109896867 malate dehydrogenase, NAD-dependent [Pseudoalteromonas atlantica T6c] 79 Q87SU7 48428249 Malate dehydrogenase 80 ABD77291 89574131 mitochondrial malate dehydrogenase 2, NAD [Aotus trivirgatus] 81 ZP_007 . . . 75854582 COG0039: Malate/lactate dehydrogenases [Vibrio sp. Ex25] 82 ZP_011 . . . 88795318 malate dehydrogenase [Alteromonas macleodii ‘Deep ecotype’] 83 ZP_012 . . . 91228842 malate dehydrogenase [Vibrio alginolyticus 12G01] 84 ZP_011 . . . 89075547 malate dehydrogenase [Photobacterium sp. SKA34] 85 ZP_012 . . . 90580811 malate dehydrogenase [Vibrio angustum S14] 86 YP_671200 110643470 malate dehydrogenase [Escherichia coli 536] 87 ABD77295 89574139 mitochondrial malate dehydrogenase 2, NAD [Equus caballus] 88 Q5E875 66774139 Malate dehydrogenase 89 ZP_007 . . . 75209673 COG0039: Malate/lactate dehydrogenases [Escherichia coli B171] 90 ZP_009 . . . 84394039 malate dehydrogenase [Vibrio splendidus 12B01] 91 ZP_007 . . . 75228638 COG0039: Malate/lactate dehydrogenases [Escherichia coli B7A] 92 ZP_010 . . . 86147822 malate dehydrogenase [Vibrio sp. MED222] 93 BAD30064 50508045 malate dehydrogenase [Moritella sp. 36B1] 94 Q4QL89 73621201 Malate dehydrogenase 95 YP_404893 82778544 malate dehydrogenase [Shigella dysenteriae Sd197] 96 BAD30059 50508035 malate dehydrogenase [Moritella sp. 47A1] 97 BAD30066 50508049 malate dehydrogenase [Moritella sp. 36G1] 98 ZP_008 . . . 77958772 COG0039: Malate/lactate dehydrogenases [Yersinia bercovieri ATCC 43970] 99 BAD30071 50508059 malate dehydrogenase [Moritella sp. 38F1] 100 ZP_008 . . . 77973999 COG0039: Malate/lactate dehydrogenases [Yersinia frederiksenii ATCC 33641] 101 ZP_008 . . . 77977600 COG0039: Malate/lactate dehydrogenases [Yersinia intermedia ATCC 29909] 102 P48364 1346511 Malate dehydrogenase 103 BAD30062 50508041 malate dehydrogenase [Moritella sp. 16H2] 104 BAD30069 50508055 malate dehydrogenase [Moritella sp. 56A1] 105 ZP_008 . . . 77961850 COG0039: Malate/lactate dehydrogenases [Yersinia mollaretii ATCC 43969] 106 ZP_012 . . . 90411956 putative malate dehydrogenase [Photobacterium profundum 3TCK] 107 Q6AW21 57012891 Malate dehydrogenase 108 Q57JA9 68052390 Malate dehydrogenase 109 ZP_012 . . . 90407531 malate dehydrogenase [Psychromonas sp. CNPT3] 110 BAD36746 51172588 malate dehydrogenase [Moritella yayanosii] 111 Q6AW23 57012892 Malate dehydrogenase 112 ABG22106 108863015 Malate dehydrogenase, glyoxysomal precursor, putative, expressed [Oryza sativa (japonica cultivar-group)] 113 ZP_005 . . . 68546610 Malate dehydrogenase, NAD-dependent, eukaryotes and gamma proteobacteria [Shewanella amazonensis SB2B] 114 BAD30060 50508037 malate dehydrogenase [Moritella sp. 47B1] 115 EAL31008 54642259 GA10540-PA [Drosophila pseudoobscura] 116 YP_736674 114046124 malate dehydrogenase, NAD-dependent [Shewanella sp. MR- 7] 117 CAJ07716 68129179 malate dehydrogenase, putative [Leishmania major] 118 ZP_009 . . . 82743617 Malate dehydrogenase, NAD-dependent, eukaryotes and gamma proteobacteria [Shewanella sp. W3-18-1] 119 Q47VL0 83288301 Malate dehydrogenase 120 ZP_011 . . . 88859032 malate dehydrogenase [Pseudoalteromonas tunicata D2] 121 YP_561838 91792187 malate dehydrogenase, NAD-dependent [Shewanella denitrificans OS217] 122 ZP_005 . . . 68544347 Malate dehydrogenase, NAD-dependent, eukaryotes and gamma proteobacteria [Shewanella baltica OS155] 123 ZP_007 . . . 75429373 malate dehydrogenase [Actinobacillus succinogenes 130Z] 124 AAW29940 56788316 malate dehydrogenase [Pasteurella trehalosi] 125 BAD30063 50508043 malate dehydrogenase [Shewanella sp. T4609] 126 EAL31009 54642260 GA10541-PA [Drosophila pseudoobscura] 127 ZP_006 . . . 69950735 Malate dehydrogenase, NAD-dependent, eukaryotes and gamma proteobacteria [Shewanella frigidimarina NCIMB 400] 128 YP_341148 77361573 malate dehydrogenase [Pseudoalteromonas haloplanktis TAC125] 129 ZP_013 . . . 106882260 malate dehydrogenase, NAD-dependent [Psychromonas ingrahamii 37] 130 BAD30068 50508053 malate dehydrogenase [Shewanella sp. 33H2] 131 ZP_008 . . . 78368581 Malate dehydrogenase, NAD-dependent, eukaryotes and gamma proteobacteria [Shewanella sp. PV-4] 132 XP_851105 74004024 PREDICTED: similar to malate dehydrogenase, mitochondrial [Canis familiaris] 133 ABD77300 89574149 mitochondrial malate dehydrogenase 2, NAD [Hippopotamus amphibius] 134 XP_816948 71656813 glycosomal malate dehydrogenase [Trypanosoma cruzi strain CL Brener] 135 CAJ19596 82655060 malate dehydrogenase [Klebsiella pneumoniae] 136 AAW29939 56788314 malate dehydrogenase [Mannheimia glucosida] 137 CAJ19583 82655034 malate dehydrogenase [Klebsiella pneumoniae] 138 CAJ19589 82655046 malate dehydrogenase [Klebsiella pneumoniae] 139 CAJ19580 82655028 malate dehydrogenase [Klebsiella pneumoniae] 140 CAJ19591 82655050 malate dehydrogenase [Klebsiella pneumoniae] 141 CAJ19595 82655058 malate dehydrogenase [Klebsiella pneumoniae] 142 CAJ19593 82655054 malate dehydrogenase [Klebsiella pneumoniae] 143 AAW29937 56788310 malate dehydrogenase [Mannheimia haemolytica] 144 XP_812467 71423452 glycosomal malate dehydrogenase [Trypanosoma cruzi strain CL Brener] 145 AAW29938 56788312 malate dehydrogenase [Mannheimia haemolytica] 146 AAW29936 56788308 malate dehydrogenase [Mannheimia haemolytica] 147 XP_849858 73993436 PREDICTED: similar to malate dehydrogenase, mitochondrial [Canis familiaris] 148 AAW28048 56718658 malate dehydrogenase [Vibrio cholerae] 149 AAU88998 52856248 malate dehydrogenase [Shigella flexneri] 150 AAW28041 56718644 malate dehydrogenase [Vibrio cholerae] 151 AAW28047 56718656 malate dehydrogenase [Vibrio mimicus] 152 AAW28045 56718652 malate dehydrogenase [Vibrio mimicus] 153 ABD77282 89574113 mitochondrial malate dehydrogenase 2, NAD [Tamandua tetradactyla] 154 AAU89018 52856288 malate dehydrogenase [Shigella boydii] 155 AAU89003 52856258 malate dehydrogenase [Escherichia coli] 156 AAU88960 52856172 malate dehydrogenase [Escherichia coli] 157 AAU88962 52856176 malate dehydrogenase [Shigella flexneri] 158 AAU88996 52856244 malate dehydrogenase [Escherichia coli] 159 AAU88972 52856196 malate dehydrogenase [Shigella boydii] 160 AAU88969 52856190 malate dehydrogenase [Shigella boydii] 161 ZP_007 . . . 75822170 COG0039: Malate/lactate dehydrogenases [Vibrio cholerae RC385] 162 AAU89062 52856376 malate dehydrogenase [Shigella boydii] 163 AAU88982 52856216 malate dehydrogenase [Shigella dysenteriae] 164 AAU88995 52856242 malate dehydrogenase [Escherichia coli] 165 AAU88993 52856238 malate dehydrogenase [Escherichia coli] 166 AAU89044 52856340 malate dehydrogenase [Shigella flexneri] 167 AAU89015 52856282 malate dehydrogenase [Shigella boydii] 168 AAU88971 52856194 malate dehydrogenase [Shigella sonnei] 169 AAW28034 56718629 malate dehydrogenase [Vibrio cholerae] 170 AAU89011 52856274 malate dehydrogenase [Shigella flexneri] 171 AAU88958 52856168 malate dehydrogenase [Escherichia coli] 172 AAU88976 52856204 malate dehydrogenase [Shigella dysenteriae] 173 AAU88991 52856234 malate dehydrogenase [Escherichia coli] 174 XP_850818 73988067 PREDICTED: similar to mitochondrial malate dehydrogenase precursor [Canis familiaris] 175 AAU86550 52697626 malate dehydrogenase [Shigella boydii] 176 AAU88979 52856210 malate dehydrogenase [Shigella dysenteriae] 177 AAU89005 52856262 malate dehydrogenase [Escherichia coli] 178 AAU89019 52856290 malate dehydrogenase [Shigella dysenteriae] 179 AAU88983 52856218 malate dehydrogenase [Shigella dysenteriae] 180 AAU86549 52697624 malate dehydrogenase [Escherichia albertii] 181 AAX85842 62532910 Mdh [Escherichia coli] 182 AAU89007 52856266 malate dehydrogenase [Shigella flexneri] 183 AAU88980 52856212 malate dehydrogenase [Shigella flexneri] 184 AAU89000 52856252 malate dehydrogenase [Shigella flexneri] 185 AAU88968 52856188 malate dehydrogenase [Shigella boydii] 186 AAU88988 52856228 malate dehydrogenase [Escherichia coli] 187 AAX85856 62532938 Mdh [Escherichia coli] 188 AAU86566 52697658 malate dehydrogenase [Shigella boydii] 189 AAU86571 52697668 malate dehydrogenase [Escherichia albertii] 190 ABF20150 94468239 Mdh [Escherichia coli] 191 AAU89051 52856354 malate dehydrogenase [Shigella boydii] 192 AAU89059 52856370 malate dehydrogenase [Shigella boydii] 193 AAU89057 52856366 malate dehydrogenase [Shigella boydii] 194 AAU89036 52856324 malate dehydrogenase [Shigella flexneri] 195 AAU86559 52697644 malate dehydrogenase [Shigella boydii] 196 ABF20151 94468241 Mdh [Escherichia coli] 197 ABF20142 94468223 Mdh [Escherichia coli] 198 AAU88990 52856232 malate dehydrogenase [Escherichia coli] 199 AAU86567 52697660 malate dehydrogenase [Shigella boydii] 200 AAU89032 52856316 malate dehydrogenase [Shigella dysenteriae] 201 AAU89012 52856276 malate dehydrogenase [Shigella flexneri]

TABLE 45 Examples of glucose-6-phosphate dehydrogenase polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_504275 50553728 hypothetical protein [Yarrowia lipolytica] 2 XP_365081 39965237 hypothetical protein MG09926.4 [Magnaporthe grisea 70-15] 3 XP_381455 46108794 G6PD_ASPNG Glucose-6-phosphate 1-dehydrogenase (G6PD) [Gibberella zeae PH-1] 4 XP_660585 67525047 glucose-6-phosphate 1-dehydrogenase [Aspergillus nidulans FGSC A4] 5 P48826 1346070 Glucose-6-phosphate 1-dehydrogenase (G6PD) 6 CAA54840 1523782 glucose-6-phosphate 1-dehydrogenase [Aspergillus niger] 7 EAU38380 114196680 glucose-6-phosphate 1-dehydrogenase [Aspergillus terreus NIH2624] 8 EAT83608 111062488 predicted protein [Phaeosphaeria nodorum SN15] 9 EAS28915 90299284 glucose-6-phosphate 1-dehydrogenase [Coccidioides immitis RS] 10 XP_754767 71000100 glucose-6-phosphate 1-dehydrogenase [Aspergillus fumigatus Af293] 11 XP_958320 85090234 GLUCOSE-6-PHOSPHATE 1-DEHYDROGENASE (G6PD) [Neurospora crassa OR74A] 12 EAQ93370 88185902 glucose-6-phosphate 1-dehydrogenase [Chaetomium globosum CBS 148.51] 13 CAA58825 642160 unnamed protein product [Emericella nidulans] 14 XP_458129 50419213 hypothetical protein DEHA0C11286g [Debaryomyces hansenii CBS767] 15 AAB25541 299248 glucose-6-phosphate dehydrogenase [Pichia jadinii = yeast, Peptide, 495 aa] 16 XP_723251 68465104 putative glucose-6-phosphate dehydrogenase [Candida albicans SC5314] 17 XP_723440 68464725 putative glucose-6-phosphate dehydrogenase [Candida albicans SC5314] 18 XP_453944 50307901 G6PD_KLULA [Kluyveromyces lactis] 19 NP_982741 45185024 ABL206Cp [Eremothecium gossypii] 20 XP_448038 50291211 unnamed protein product [Candida glabrata] 21 XP_572045 58269778 glucose-6-phosphate 1-dehydrogenase [Cryptococcus neoformans var. neoformans JEC21] 22 NP_014158 6324088 Glucose-6-phosphate dehydrogenase (G6PD), catalyzes the first step of the pentose phosphate pathway; involved in adapting to oxidatve stress; homolog of the human G6PD which is deficient in patients with hemolytic anemia; Zwf1p [Saccharomyces cerevisiae] 23 AAT93017 51013447 YNL241C [Saccharomyces cerevisiae] 24 AAA34619 171545 glucose-6-phosphate dehydrogenase (ZWF1) (EC 1.1.1.49) 25 XP_761077 71021693 hypothetical protein UM04930.1 [Ustilago maydis 521] 26 NP_593344 63054535 glucose-6-phosphate 1-dehydrogenase [Schizosaccharomyces pombe 972h-] 27 XP_644814 66822919 glucose 6-phosphate-1-dehydrogenase [Dictyostelium discoideum AX4] 28 ABD72519 89357348 glucose 6-phosphate dehydrogenase [Trypanosoma cruzi] 29 BAE99888 110735823 glucose-6-phosphate dehydrogenase [Arabidopsis thaliana] 30 CAB52675 5732197 glucose-6-phosphate 1-dehydrogenase [Arabidopsis thaliana] 31 CAB08746 2104434 SPAC3A12.18 [Schizosaccharomyces pombe] 32 ABD72517 89357344 glucose 6-phosphate dehydrogenase [Trypanosoma cruzi] 33 ABD72518 89357346 glucose 6-phosphate dehydrogenase [Trypanosoma cruzi] 34 BAB96757 21262179 glucose-6-phosphate dehydrogenase 1 [Chlorella vulgaris] 35 XP_822502 71746894 glucose-6-phosphate 1-dehydrogenase [Trypanosoma brucei TREU927] 36 CAJ07708 68129171 glucose-6-phosphate dehydrogenase [Leishmania major] 37 CAC07816 10045209 glucose-6-phosphate 1-dehydrogenase [Trypanosoma brucei] 38 BAF15046 113564703 Os04g0485300 [Oryza sativa (japonica cultivar-group)] 39 AAO37825 28261397 glucose-6-phosphate dehydrogenase [Leishmania mexicana] 40 AAM64228 27434608 glucose-6-phosphate dehydrogenase [Leishmania mexicana amazonensis] 41 XP_001 . . . 110750934 PREDICTED: similar to Zwischenferment CG12529-PA, isoform A [Apis mellifera] 42 XP_974096 91082561 PREDICTED: similar to CG12529-PA, isoform A [Tribolium castaneum] 43 ABE83959 92876243 Glucose-6-phosphate dehydrogenase [Medicago truncatula] 44 BAE38077 74138546 unnamed protein product [Mus musculus] 45 NP_001 . . . 79313844 G6PD5; glucose-6-phosphate 1-dehydrogenase [Arabidopsis thaliana] 46 CAB52674 5732195 glucose-6-phosphate 1-dehydrogenase [Arabidopsis thaliana] 47 AAD11426 4206114 cytoplasmic glucose-6-phosphate 1-dehydrogenase [Mesembryanthemum crystallinum] 48 Q42919 3023815 Glucose-6-phosphate 1-dehydrogenase, cytoplasmic isoform (G6PD) 49 AAL79959 19071787 glucose-6-phosphate dehydrogenase [Oryza sativa (japonica cultivar-group)] 50 CAE62054 39593761 Hypothetical protein CBG06072 [Caenorhabditis briggsae] 51 EAL31619 54642875 GA11679-PA [Drosophila pseudoobscura] 52 Q00612 54037161 Glucose-6-phosphate 1-dehydrogenase X (G6PD) 53 AAH75663 49523350 Glucose-6-phosphate dehydrogenase X-linked [Mus musculus] 54 AAB69319 2352923 cytosolic glucose-6-phosphate dehydrogenase 2 [Petroselinum crispum] 55 AAM64230 27434612 glucose-6-phosphate dehydrogenase [Leishmania guyanensis] 56 ABB29564 78183137 putative Zwischenferment [Drosophila yakuba] 57 AAL57688 18086470 AT3g27300/K17E12_12 [Arabidopsis thaliana] 58 NP_728287 24643352 Zwischenferment CG12529-PB, isoform B [Drosophila melanogaster] 59 EAT38627 108874402 glucose-6-phosphate 1-dehydrogenase [Aedes aegypti] 60 AAR26303 89214190 glucose-6-phosphate dehydrogenase [Populus suaveolens] 61 AAA99073 1304700 glucose-6-phosphate 1-dehydrogenase [Drosophila melanogaster] 62 AAK93503 15292469 SD03244p [Drosophila melanogaster] 63 AAB02809 1304686 glucose-6-phosphate 1-dehydrogenase [Drosophila melanogaster] 64 NP_523411 24643350 Zwischenferment CG12529-PA, isoform A [Drosophila melanogaster] 65 AAA99107 1305086 glucose-6-phosphate 1-dehydrogenase [Drosophila melanogaster] 66 ABB29562 78183133 putative Zwischenferment [Drosophila simulans] 67 NP_502129 17538218 B0035.5 [Caenorhabditis elegans] 68 AAZ23850 71089991 glucose-6-phosphate dehydrogenase [Boophilus microplus] 69 P37830 585165 Glucose-6-phosphate 1-dehydrogenase, cytoplasmic isoform (G6PD) 70 AAB02812 1304692 glucose-6-phosphate 1-dehydrogenase [Drosophila melanogaster] 71 ABB55386 81075965 glucose-6-phosphate 1-dehydrogenase cytoplasmic isoform- like [Solanum tuberosum] 72 CAJ83683 89273424 glucose-6-phosphate dehydrogenase [Xenopus tropicalis] 73 NP_001 . . . 62859893 glucose-6-phosphate dehydrogenase 2 [Xenopus tropicalis] 74 ABB29563 78183135 putative Zwischenferment [Drosophila teissieri] 75 AAX45785 61394184 glucose-6-phosphate dehydrogenase isoform B [Ips typographus] 76 AAX45784 61394183 glucose-6-phosphate dehydrogenase isoform A [Ips typographus] 77 AAH81820 51980296 Glucose-6-phosphate dehydrogenase X-linked [Rattus norvegicus] 78 XP_311452 58381765 ENSANGP00000018551 [Anopheles gambiae str. PEST] 79 XP_538209 74009187 PREDICTED: similar to Glucose-6-phosphate 1- dehydrogenase (G6PD) [Canis familiaris] 80 AAH59324 111185531 Unknown (protein for MGC: 69058) [Xenopus laevis] 81 ABB29560 78183129 putative Zwischenferment [Drosophila erecta] 82 Q27638 3023810 Glucose-6-phosphate 1-dehydrogenase (G6PD) 83 Q29492 2494652 Glucose-6-phosphate 1-dehydrogenase (G6PD) 84 BAF09259 113536876 Os02g0600400 [Oryza sativa (japonica cultivar-group)] 85 O55044 62510568 Glucose-6-phosphate 1-dehydrogenase (G6PD) 86 CAA04993 3021510 glucose-6-phosphate dehydrogenase [Nicotiana tabacum] 87 ABB29561 78183131 putative Zwischenferment [Drosophila orena] 88 BAA97662 8918502 glucose-6-phosphate dehydrogenase [Triticum aestivum] 89 BAD17920 46849421 glucose-6-phosphate 1-dehydrogenase [Acipenser baerii] 90 XP_699168 68440567 PREDICTED: similar to glucose-6-phosphate dehydrogenase [Danio rerio] 91 AAB96363 2734869 glucose-6-phosphate dehydrogenase [Takifugu rubripes] 92 NP_000393 109389365 glucose-6-phosphate dehydrogenase isoform a [Homo sapiens] 93 NP_001 . . . 108773793 glucose-6-phosphate dehydrogenase isoform b [Homo sapiens] 94 AAA52500 182871 glucose-6-phosphate dehydrogenase variant A-(EC 1.1.1.49) 95 P11413 120731 Glucose-6-phosphate 1-dehydrogenase (G6PD) 96 AAA92653 1203978 G6PD [Homo sapiens] 97 AAX43335 61369430 glucose-6-phosphate dehydrogenase [synthetic construct] 98 P41571 1169799 Glucose-6-phosphate 1-dehydrogenase (G6PD) (Zwischenferment) 99 2BHLB 66361518 Chain B, X-Ray Structure Of Human Glucose-6-Phosphate Dehydrogenase (Deletion Variant) Complexed With Glucose- 6-Phosphate 100 ABC74527 85700174 glucose-6-phosphate dehydrogenase [Populus trichocarpa] 101 ABC74528 85700176 glucose-6-phosphate dehydrogenase [Populus trichocarpa] 102 AAA51463 157470 glucose-6-phosphate dehydrogenase 103 XP_001 . . . 109132852 PREDICTED: glucose-6-phosphate dehydrogenase isoform 1 [Macaca mulatta] 104 1QKIH 7546530 Chain H, X-Ray Structure Of Human Glucose 6-Phosphate Dehydrogenase (Variant Canton R4591) Complexed With Structural Nadp+ 105 XP_001 . . . 109132850 PREDICTED: glucose-6-phosphate dehydrogenase isoform 2 [Macaca mulatta] 106 BAA97663 8918504 glucose-6-phosphate dehydrogenase [Triticum aestivum] 107 CAA04992 3021508 glucose-6-phosphate dehydrogenase [Nicotiana tabacum] 108 AAB69318 2352921 cytosolic glucose-6-phosphate dehydrogenase 1 [Petroselinum crispum] 109 XP_307095 58375370 ENSANGP00000012074 [Anopheles gambiae str. PEST] 110 AAA41179 204197 glucose-6-phosphate dehydrogenase 111 AAA63175 182890 glucose-6-phosphate dehydrogenase [Homo sapiens] 112 BAA97664 8918506 glucose-6-phosphate dehydrogenase [Triticum aestivum] 113 AAW24823 56753227 SJCHGC02527 protein [Schistosoma japonicum] 114 CAA58590 5459313 glucose-6-phosphate 1-dehydrogenase [Takifugu rubripes] 115 CAG07451 47228719 unnamed protein product [Tetraodon nigroviridis] 116 CAJ28912 113207852 glucose-6-phosphate 1-dehydrogenase [Crassostrea gigas] 117 ABC25881 83628088 glucose-6-phosphate 1-dehydrogenase [Homo sapiens] 118 ABC25981 83628208 glucose-6-phosphate 1-dehydrogenase [Homo sapiens] 119 ABC25826 83628022 glucose-6-phosphate 1-dehydrogenase [Homo sapiens] 120 BAD17912 46849405 glucose-6-phosphate 1-dehydrogenase [Amia calva] 121 BAD17877 46849335 glucose-6-phosphate 1-dehydrogenase [Protopterus annectens] 122 BAD17947 46849475 glucose-6-phosphate 1-dehydrogenase [Callorhinchus callorynchus] 123 AAI20828 111306848 Glucose-6-phosphate dehydrogenase 2 [Mus musculus] 124 BAD17927 46849435 glucose-6-phosphate 1-dehydrogenase [Polypterus ornatipinnis] 125 BAD17898 46849377 glucose-6-phosphate 1-dehydrogenase [Oryzias latipes] 126 BAD17891 46849363 glucose-6-phosphate 1-dehydrogenase [Ambystoma mexicanum] 127 BAD17951 46849483 glucose-6-phosphate 1-dehydrogenase [Lethenteron reissneri] 128 AAF19030 7629275 glucose-6-phosphate-1-dehydrogenase; G6PD [Pimephales promelas] 129 BAD17941 46849463 glucose-6-phosphate 1-dehydrogenase [Potamotrygon motoro] 130 BAD17954 46849489 glucose-6-phosphate 1-dehydrogenase [Branchiostoma belcheri] 131 BAD17905 46849391 glucose-6-phosphate 1-dehydrogenase [Lepisosteus osseus] 132 BAD17884 46849349 glucose-6-phosphate 1-dehydrogenase [Lepidosiren paradoxa] 133 BAD17934 46849449 glucose-6-phosphate 1-dehydrogenase [Cephaloscyllium umbratile] 134 ABC74529 85700178 glucose-6-phosphate dehydrogenase [Populus trichocarpa] 135 CAB52685 5734379 plastidic glucose-6-phosphate dehydrogenase [Dunaliella bioculata] 136 Q43793 3023817 Glucose-6-phosphate 1-dehydrogenase, chloroplast precursor (G6PD) 137 AAF87216 9392607 plastidic glucose 6-phosphate dehydrogenase [Nicotiana tabacum] 138 BAC23041 24745908 glucose 6-phosphate dehydrogenase [Solanum tuberosum] 139 CAB52708 5734502 glucose-6-phosphate 1-dehydrogenase [Solanum tuberosum] 140 BAF21344 113610966 Os07g0406300 [Oryza sativa (japonica cultivar-group)] 141 AAQ02671 33304517 putative plastidic glucose-6-phosphate dehydrogenase [Oryza sativa (japonica cultivar-group)] 142 AAW82643 58803037 hepatic glucose-6-phosphate dehydrogenase [Rhabdosargus sarba] 143 BAF12270 113548827 Os03g0412800 [Oryza sativa (japonica cultivar-group)] 144 AAN72144 25083966 glucose-6-phosphate dehydrogenase [Arabidopsis thaliana] 145 AAB69317 2352919 plastidic glucose-6-phosphate dehydrogenase [Petroselinum crispum] 146 O24357 3334193 Glucose-6-phosphate 1-dehydrogenase, chloroplast precursor (G6PD) 147 AAO23597 27764952 At1g24280/F3I6_22 [Arabidopsis thaliana] 148 Q8L743 25452975 Glucose-6-phosphate 1-dehydrogenase 3, chloroplast precursor (G6PD3) (G6PDH3) 149 CAA59011 1166405 glucose-6-phosphate 1-dehydrogenase [Arabidopsis thaliana] 150 CAA59012 1174336 glucose-6-phosphate 1-dehydrogenase [Arabidopsis thaliana] 151 CAC05439 9955555 glucose-6-phosphate 1-dehydrogenase [Arabidopsis thaliana] 152 Q9FY99 25452980 Glucose-6-phosphate 1-dehydrogenase 2, chloroplast precursor (G6PD2) (G6PDH2) 153 AAL57678 18086448 AT5g13110/T19L5_70 [Arabidopsis thaliana] 154 Q43839 3023818 Glucose-6-phosphate 1-dehydrogenase, chloroplast precursor (G6PD) 155 CAA04696 3021305 plastidic glucose-6-phosphate dehydrogenase [Arabidopsis thaliana] 156 CAB52681 5734372 glucose-6-phosphate 1-dehydrogenase [Cyanidium caldarium] 157 CAA04994 3021532 glucose-6-phosphate dehydrogenase [Nicotiana tabacum] 158 Q7YS37 62510624 Glucose-6-phosphate 1-dehydrogenase (G6PD) 159 CAG04059 47229307 unnamed protein product [Tetraodon nigroviridis] 160 CAD28862 29149997 glucose 6 phosphate dehydrogenase [Acraea encedana] 161 CAD28863 29149999 glucose 6 phosphate dehydrogenase [Acraea encedana] 162 AAM64231 27434614 glucose-6-phosphate dehydrogenase [Leishmania braziliensis] 163 AAG28728 11066848 glucose-6-phosphate-dehydrogenase [Drosophila mauritiana] 164 AAG28730 11066852 glucose-6-phosphate-dehydrogenase [Drosophila sechellia] 165 ABF96583 108708788 Glucose-6-phosphate 1-dehydrogenase, chloroplast precursor, putative, expressed [Oryza sativa (japonica cultivar-group)] 166 CAA03941 2276348 Glucose-6-phosphate dehydrogenase [Spinacia oleracea] 167 XP_583628 76658972 PREDICTED: similar to Glucose-6-phosphate 1- dehydrogenase (G6PD) [Bos taurus] 168 AAA52499 182869 glucose-6-phosphate dehydrogenase 169 AAM64229 27434610 glucose-6-phosphate dehydrogenase [Leishmania mexicana] 170 NP_587749 19075249 hypothetical protein SPCC794.01c [Schizosaccharomyces pombe 972h-] 171 AAR12945 38156652 glucose-6-phosphate dehydrogenase [Drosophila mojavensis] 172 AAR12953 38156668 glucose-6-phosphate dehydrogenase [Drosophila mojavensis] 173 AAR12946 38156654 glucose-6-phosphate dehydrogenase [Drosophila mojavensis] 174 AAR12943 38156648 glucose-6-phosphate dehydrogenase [Drosophila mojavensis] 175 AAR12952 38156666 glucose-6-phosphate dehydrogenase [Drosophila mojavensis] 176 BAF11858 113548415 Os03g0318500 [Oryza sativa (japonica cultivar-group)] 177 XP_559252 57942974 ENSANGP00000028421 [Anopheles gambiae str. PEST] 178 NP_593614 19114526 hypothetical protein SPAC3C7.13c [Schizosaccharomyces pombe 972h-] 179 BAD94743 62321397 glucose-6-phosphate dehydrogenase [Arabidopsis thaliana] 180 CAJ20381 95007160 glucose-6-phosphate-1-dehydrogenase [Toxoplasma gondii] 181 YP_605437 94986073 glucose-6-phosphate 1-dehydrogenase [Deinococcus geothermalis DSM 11300] 182 CAE51228 37651949 glucose 6 phosphate dehydrogenase [Adalia bipunctata] 183 YP_007820 46446455 putative glucose-6-phosphate [Candidatus Protochlamydia amoebophila UWE25] 184 NP_295319 15806604 glucose-6-phosphate 1-dehydrogenase [Deinococcus radiodurans R1] 185 CAE51222 37651937 glucose 6 phosphate dehydrogenase [Adalia bipunctata] 186 CAE51229 37651951 glucose 6 phosphate dehydrogenase [Adalia decempunctata] 187 ZP_007 . . . 76260057 Glucose-6-phosphate dehydrogenase [Chloroflexus aurantiacus J-10-fl] 188 ZP_002 . . . 47096622 glucose-6-phosphate 1-dehydrogenase [Listeria monocytogenes str. 1/2a F6854] 189 ZP_004 . . . 66965919 Glucose-6-phosphate dehydrogenase [Arthrobacter sp.FB24] 190 ZP_002 . . . 47093301 glucose-6-phosphate 1-dehydrogenase [Listeria monocytogenes str. 4b H7858] 191 NP_471419 16801151 hypothetical protein lin2085 [Listeria innocua Clip11262] 192 NP_693860 23100393 glucose-6-phosphate 1-dehydrogenase [Oceanobacillus iheyensis HTE831] 193 NP_829405 29840299 glucose-6-phosphate 1-dehydrogenase [Chlamydophila caviae GPIC] 194 CAJ71041 91203388 strongly similar to glucose-6-phosphate dehydrogenase [Candidatus Kuenenia stuttgartiensis] 195 YP_629214 108761123 glucose-6-phosphate 1-dehydrogenase [Myxococcus xanthus DK 1622] 196 ZP_009 . . . 84387050 glucose-6-phosphate 1-dehydrogenase [Vibrio splendidus 12B01] 197 ZP_011 . . . 88856188 glucose-6-phosphate 1-dehydrogenase [marine actinobacterium PHSC20C1] 198 NP_827489 29832855 glucose-6-phosphate 1-dehydrogenase [Streptomyces avermitilis MA-4680] 199 CAH10104 88319768 putative glucose-6-phosphate-1-dehydrogenase [Streptomyces sp. SCC 2136] 200 YP_148187 56420869 glucose-6-phosphate 1-dehydrogenase [Geobacillus kaustophilus HTA426] 201 ZP_006 . . . 71368421 Glucose-6-phosphate dehydrogenase [Nocardioides sp. JS614]

TABLE 46 Examples of 6-phosphogluconate dehydrogenase polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_500938 50546937 hypothetical protein [Yarrowia lipolytica]. 2 XP_458726 50420381 hypothetical protein DEHA0D06820g [Debaryomyces hansenii CBS767] 3 XP_451408 50302941 unnamed protein product [Kluyveromyces lactis] 4 O13287 3334110 6-phosphogluconate dehydrogenase, decarboxylating 5 XP_722227 68467588 hypothetical protein CaO19_5024 [Candida albicans SC5314] 6 NP_012053 6321977 6-phosphogluconate dehydrogenase (decarboxylating), catalyzes an NADPH regenerating reaction in the pentose phosphate pathway; required for growth on D-glucono-delta- lactone and adaptation to oxidative stress; Gnd1p [Saccharomyces cerevisiae] 7 AAO32396 28564035 GND1 [Saccharomyces bayanus] 8 XP_750696 70991695 6-phosphogluconate dehydrogenase, decarboxylating [Aspergillus fumigatus Af293] 9 CAD80254 29409963 6-phosphogluconate dehydrogenase [Aspergillus niger] 10 XP_661558 67526993 hypothetical protein AN3954.2 [Aspergillus nidulans FGSC A4] 11 AAO32456 28564155 GND1 [Saccharomyces servazzii] 12 AAO32497 28564385 GND1 [Saccharomyces castellii] 13 BAE57349 83767210 unnamed protein product [Aspergillus oryzae] 14 AAT92830 51013073 YGR256W [Saccharomyces cerevisiae] 15 XP_449923 50295024 hypothetical protein CAGL0M13343g [Candida glabrata CBS138] 16 NP_985676 45198647 AFR129Wp [Eremothecium gossypii] 17 EAS35799 90306168 6-phosphogluconate dehydrogenase, decarboxylating [Coccidioides immitis RS] 18 XP_758724 71014537 hypothetical protein UM02577.1 [Ustilago maydis 521] 19 O60037 12229635 6-phosphogluconate dehydrogenase, decarboxylating 20 XP_964959 85115938 hypothetical protein [Neurospora crassa OR74A] 21 XP_381287 46108458 hypothetical protein FG01111.1 [Gibberella zeae PH-1] 22 EAT76717 111055597 predicted protein [Phaeosphaeria nodorum SN15] 23 XP_369069 39975357 hypothetical protein MG00175.4 [Magnaporthe grisea 70-15] 24 XP_567793 58260766 phosphogluconate dehydrogenase (decarboxylating) [Cryptococcus neoformans var. neoformans JEC21] 25 NP_595095 19111887 6-phosphogluconate dehydrogenase [Schizosaccharomyces pombe 972h-] 26 EAQ92608 88185140 hypothetical protein CHGG_00843 [Chaetomium globosum CBS 148.51] 27 T42523 11251352 probable phosphogluconate dehydrogenase (decarboxylating) (EC 1.1.1.44) - fission yeast (Schizosaccharomyces pombe ) (fragment) 28 XP_781394 72005545 PREDICTED: similar to 6-phosphogluconate dehydrogenase, decarboxylating [Strongylocentrotus purpuratus] 29 BAD98151 63003720 6-phosphogluconate dehydrogenase [Ascidia sydneiensis samea] 30 XP_625090 66547531 PREDICTED: similar to 6-phosphogluconate dehydrogenase, decarboxylating, partial [Apis mellifera] 31 CAG32303 53134115 hypothetical protein [Gallus gallus] 32 2PGD 999886 Chain, 6-Phosphogluconate Dehydrogenase (6-Pgdh) (E.C.1.1.1.44) 33 NP_001 . . . 57164179 6-phosphogluconate dehydrogenase (decarboxylating) [Ovis aries] 34 XP_911640 82896232 PREDICTED: similar to 6-phosphogluconate dehydrogenase, decarboxylating isoform 6 [Mus musculus] 35 BAE31577 74225286 unnamed protein product [Mus musculus] 36 AAX43359 61369608 phosphogluconate dehydrogenase [synthetic construct] 37 CAI95751 66347393 phosphogluconate dehydrogenase [Homo sapiens] 38 XP_993478 94374014 PREDICTED: similar to 6-phosphogluconate dehydrogenase, decarboxylating isoform 4 [Mus musculus] 39 AAH11329 15030150 Phosphogluconate dehydrogenase [Mus musculus] 40 AAI02179 73586529 LOC514939 protein [Bos taurus] 41 AAA75302 984325 phosphogluconate dehydrogenase [Homo sapiens] 42 AAH59958 37747616 MGC68486 protein [Xenopus laevis] 43 ZP_005 . . . 67931108 6-phosphogluconate dehydrogenase, decarboxylating [Solibacter usitatus Ellin6076] 44 EAT42557 108878332 6-phosphogluconate dehydrogenase [Aedes aegypti] 45 XP_535411 73950940 PREDICTED: similar to 6-phosphogluconate dehydrogenase, decarboxylating isoform 1 [Canis familiaris] 46 XP_313091 58384192 ENSANGP00000012857 [Anopheles gambiae str. PEST] 47 CAG07546 47228814 unnamed protein product [Tetraodon nigroviridis] 48 YP_007316 46445951 6-phosphogluconate dehydrogenase [Candidatus Protochlamydia amoebophila UWE25] 49 NP_998717 47087205 phosphogluconate hydrogenase isoform 2 [Danio rerio] 50 XP_972051 91094851 PREDICTED: similar to phosphogluconate hydrogenase isoform 2 [Tribolium castaneum] 51 CAE70848 39594980 Hypothetical protein CBG17632 [Caenorhabditis briggsae] 52 XP_001 . . . 94372948 PREDICTED: similar to 6-phosphogluconate dehydrogenase, decarboxylating isoform 9 [Mus musculus] 53 NP_998618 47087439 phosphogluconate hydrogenase isoform 1 [Danio rerio] 54 AAH95571 63102189 Pgd protein [Danio rerio] 55 XP_993446 94374016 PREDICTED: similar to 6-phosphogluconate dehydrogenase, decarboxylating isoform 3 [Mus musculus] 56 EAL31500 54642755 GA17642-PA [Drosophila pseudoobscura] 57 Q17761 74962414 6-phosphogluconate dehydrogenase, decarboxylating 58 P41573 1168233 6-phosphogluconate dehydrogenase, decarboxylating 59 AAL90185 19528141 AT26455p [Drosophila melanogaster] 60 NP_476860 24639279 Phosphogluconate dehydrogenase CG3724-PA [Drosophila melanogaster] 61 NP_001 . . . 114053253 6-phosphogluconate dehydrogenase [Bombyx mori] 62 P41570 1168228 6-phosphogluconate dehydrogenase, decarboxylating 63 CAJ43391 95140247 6-phosphogluconate dehydrogenase [Bactrocera oleae] 64 CAJ43390 95140245 6-phosphogluconate dehydrogenase [Ceratitis capitata] 65 AAP92648 33086672 Cc2-27 [Rattus norvegicus] 66 YP_661682 109898427 6-phosphogluconate dehydrogenase, decarboxylating [Pseudoalteromonas atlantica T6c] 67 XP_592859 76637462 PREDICTED: similar to 6-phosphogluconate dehydrogenase, decarboxylating [Bos taurus] 68 YP_526109 90020282 6-phosphogluconate dehydrogenase, decarboxylating [Saccharophagus degradans 2-40] 69 ZP_001 . . . 53733078 COG0362: 6-phosphogluconate dehydrogenase [Haemophilus influenzae R2846] 70 NP_761502 27365974 6-phosphogluconate dehydrogenase [Vibrio vulnificus CMCP6] 71 YP_391484 78485559 6-phosphogluconate dehydrogenase, decarboxylating [Thiomicrospira crunogena XCL-2] 72 YP_248262 68249150 6-phosphogluconate dehydrogenase [Haemophilus influenzae 86-028NP] 73 NP_934400 37679791 6-phosphogluconate dehydrogenase [Vibrio vulnificus YJ016] 74 ZP_007 . . . 75857693 COG0362: 6-phosphogluconate dehydrogenase [Vibrio sp. Ex25] 75 NP_798087 28898482 6-phosphogluconate dehydrogenase [Vibrio parahaemolyticus RIMD 2210633] 76 YP_206428 59713653 6-phosphogluconate dehydrogenase [Vibrio fischeri ES114] 77 ZP_001 . . . 53733322 COG0362: 6-phosphogluconate dehydrogenase [Haemophilus influenzae R2866] 78 ZP_012 . . . 91224436 6-phosphogluconate dehydrogenase [Vibrio alginolyticus 12G01] 79 AAB20377 239383 6-phosphogluconate dehydrogenase [sheep, Peptide, 466 aa] 80 ZP_010 . . . 86145883 6-phosphogluconate dehydrogenase [Vibrio sp. MED222] 81 ZP_007 . . . 75432781 Phosphogluconate dehydrogenase (decarboxylating) [Actinobacillus succinogenes 130Z] 82 ZP_001 . . . 46143450 COG0362: 6-phosphogluconate dehydrogenase [Actinobacillus pleuropneumoniae serovar 1 str. 4074] 83 ZP_012 . . . 90578957 6-phosphogluconate dehydrogenase [Vibrio angustum S14] 84 YP_087205 52424068 6-phosphogluconate dehydrogenase [Mannheimia succiniciproducens MBEL55E] 85 ZP_011 . . . 89073009 6-phosphogluconate dehydrogenase [Photobacterium sp. SKA34] 86 NP_273081 15675963 6-phosphogluconate dehydrogenase [Neisseria meningitidis MC58] 87 ZP_007 . . . 75825242 COG0362: 6-phosphogluconate dehydrogenase [Vibrio cholerae O395] 88 ZP_007 . . . 75822319 COG0362: 6-phosphogluconate dehydrogenase [Vibrio cholerae RC385] 89 ZP_007 . . . 75831146 COG0362: 6-phosphogluconate dehydrogenase [Vibrio cholerae MO10] 90 YP_719859 113461790 6-phosphogluconate dehydrogenase [Haemophilus somnus 129PT] 91 ZP_012 . . . 90411828 6-phosphogluconate dehydrogenase [Photobacterium profundum 3TCK] 92 YP_129657 54308637 6-phosphogluconate dehydrogenase [Photobacterium profundum SS9] 93 ZP_001 . . . 53728695 COG0362: 6-phosphogluconate dehydrogenase [Haemophilus somnus 2336] 94 BAD36766 51241757 6-phosphogluconate dehydrogenase [Cyanidioschyzon merolae] 95 P70718 2492494 6-phosphogluconate dehydrogenase, decarboxylating 96 XP_642122 66816225 6-phosphogluconate dehydrogenase (decarboxylating) [Dictyostelium discoideum AX4] 97 Q7VMX4 71152205 6-phosphogluconate dehydrogenase, decarboxylating 98 NP_283102 15793280 6-phosphogluconate dehydrogenase [Neisseria meningitidis Z2491] 99 NP_246493 15603419 6-phosphogluconate dehydrogenase [Pasteurella multocida subsp. multocida str. Pm70] 100 AAO19943 27573087 6-phosphogluconate dehydrogenase, decarboxylating [Neisseria gonorrhoeae] 101 AAO19944 27573089 6-phosphogluconate dehydrogenase, decarboxylating [Neisseria gonorrhoeae] 102 YP_208939 59802227 6-phosphogluconate dehydrogenase [Neisseria gonorrhoeae FA 1090] 103 AAO19934 27573069 6-phosphogluconate dehydrogenase, decarboxylating [Neisseria gonorrhoeae] 104 AAO19942 27573085 6-phosphogluconate dehydrogenase, decarboxylating [Neisseria gonorrhoeae] 105 ZP_013 . . . 106881674 6-phosphogluconate dehydrogenase, decarboxylating [Psychromonas ingrahamii 37] 106 XP_515670 55599346 PREDICTED: similar to 6-phosphogluconate dehydrogenase, decarboxylating [Pan troglodytes] 107 ZP_010 . . . 87309497 6-phosphogluconate dehydrogenase [Blastopirellula marina DSM 3645] 108 NP_865160 32472166 6-phosphogluconate dehydrogenase [Rhodopirellula baltica SH 1] 109 AAW29923 56788282 6-phosphogluconate dehydrogenase [Mannheimia haemolytica] 110 AAW29924 56788284 6-phosphogluconate dehydrogenase [Mannheimia glucosida] 111 AAW29925 56788286 6-phosphogluconate dehydrogenase [Pasteurella trehalosi] 112 NP_218771 15639322 6-phosphogluconate dehydrogenase [Treponema pallidum subsp. pallidum str. Nichols] 113 YP_148197 56420879 6-phosphogluconate dehydrogenase [Geobacillus kaustophilus HTA426] 114 YP_356348 77918533 probable phosphogluconate dehydrogenase (decarboxylating) [Pelobacter carbinolicus DSM 2380] 115 NP_390267 50812264 6-phosphogluconate dehydrogenase [Bacillus subtilis subsp. subtilis str. 168] 116 YP_092134 52786305 6-phosphogluconate dehydrogenase [Bacillus licheniformis ATCC 14580] 117 ZP_011 . . . 89099010 6-phosphogluconate dehydrogenase [Bacillus sp. NRRL B- 14911] 118 NP_691106 23097640 6-phosphogluconate dehydrogenase [Oceanobacillus iheyensis HTE831] 119 YP_219832 62185047 6-phosphogluconate dehydrogenase [Chlamydophila abortus S26/3] 120 ZP_002 . . . 47565365 6-phosphogluconate dehydrogenase, decarboxylating [Bacillus cereus G9241] 121 YP_026451 49183199 6-phosphogluconate dehydrogenase [Bacillus anthracis str. Sterne] 122 YP_034514 49476731 6-phosphogluconate dehydrogenase [Bacillus thuringiensis serovar konkukian str. 97-27] 123 YP_081773 52145056 6-phosphogluconate dehydrogenase [Bacillus cereus E33L] 124 YP_515493 89898383 6-phosphogluconate dehydrogenase [Chlamydophila felis Fe/C-56] 125 NP_814782 29375628 6-phosphogluconate dehydrogenase [Enterococcus faecalis V583] 126 ZP_003 . . . 65317607 COG0362: 6-phosphogluconate dehydrogenase [Bacillus anthracis str. A2012] 127 YP_175422 56963691 6-phosphogluconate dehydrogenase [Bacillus clausii KSM- K16] 128 ZP_006 . . . 69247351 6-phosphogluconate dehydrogenase, decarboxylating [Enterococcus faecium DO] 129 NP_810135 29346632 6-phosphogluconate dehydrogenase [Bacteroides thetaiotaomicron VPI-5482] 130 NP_829300 29840194 6-phosphogluconate dehydrogenase [Chlamydophila caviae GPIC] 131 AAL76325 18644678 6-phosphogluconate dehydrogenase [Porphyra yezoensis] 132 YP_546705 91776949 6-phosphogluconate dehydrogenase, decarboxylating [Methylobacillus flagellatus KT] 133 ZP_004 . . . 67546118 6-phosphogluconate dehydrogenase, decarboxylating [Burkholderia vietnamiensis G4] 134 Q9CHU6 18202802 6-phosphogluconate dehydrogenase, decarboxylating 135 ZP_006 . . . 74014955 6-phosphogluconate dehydrogenase, decarboxylating [Burkholderia ambifaria AMMD] 136 YP_372936 78063028 6-phosphogluconate dehydrogenase [Burkholderia sp. 383] 137 YP_624314 107026803 6-phosphogluconate dehydrogenase, decarboxylating [Burkholderia cenocepacia AU 1054] 138 NP_928851 37525507 6-phosphogluconate dehydrogenase [Photorhabdus luminescens subsp. laumondii TTO1] 139 ZP_002 . . . 47095960 6-phosphogluconate dehydrogenase, decarboxylating [Listeria monocytogenes str. 1/2a F6854] 140 YP_438830 83717630 6-phosphogluconate dehydrogenase, decarboxylating [Burkholderia thailandensis E264] 141 P96789 8134294 6-phosphogluconate dehydrogenase, decarboxylating 142 NP_924063 37520686 6-phosphogluconate dehydrogenase [Gloeobacter violaceus PCC 7421] 143 AAA24207 146938 6-phosphogluconate dehydrogenase 144 ZP_009 . . . 84360352 COG0362: 6-phosphogluconate dehydrogenase [Burkholderia dolosa AUO158] 145 YP_335987 76817886 6-phosphogluconate dehydrogenase [Burkholderia pseudomallei 1710b] 146 AAA24495 147501 6-phosphogluconate dehydrogenase 147 NP_310857 15832084 6-phosphogluconate dehydrogenase [Escherichia coli O157:H7 str. Sakai] 148 ZP_009 . . . 83571257 COG0362: 6-phosphogluconate dehydrogenase [Shigella dysenteriae 1012] 149 ZP_007 . . . 75229110 COG0362: 6-phosphogluconate dehydrogenase [Escherichia coli B7A] 150 ZP_013 . . . 100915196 hypothetical protein Bmal10_03003493 [Burkholderia mallei 10399] 151 AAG35219 11464518 6-phosphogluconate dehydrogenase [Escherichia coli] 152 ZP_013 . . . 100236192 hypothetical protein Bpse4_03000697 [Burkholderia pseudomallei 406e] 153 YP_669972 110642242 6-phosphogluconate dehydrogenase, decarboxylating [Escherichia coli 536] 154 ZP_013 . . . 100062231 hypothetical protein BpseP_03002198 [Burkholderia pseudomallei Pasteur] 155 YP_114383 53803994 6-phosphogluconate dehydrogenase [Methylococcus capsulatus str. Bath] 156 AAA24208 146940 6-phosphogluconate dehydrogenase 157 YP_310991 74312572 gluconate-6-phosphate dehydrogenase, decarboxylating [Shigella sonnei Ss046] 158 ZP_003 . . . 48870455 COG0362: 6-phosphogluconate dehydrogenase [Pediococcus pentosaceus ATCC 25745] 159 ZP_007 . . . 75514070 COG0362: 6-phosphogluconate dehydrogenase [Escherichia coli 53638] 160 YP_049550 50120383 6-phosphogluconate dehydrogenase [Erwinia carotovora subsp. atroseptica SCRI1043] 161 AP_002630 89108850 gluconate-6-phosphate dehydrogenase, decarboxylating [Escherichia coli W3110] 162 ZP_007 . . . 75259178 COG0362: 6-phosphogluconate dehydrogenase [Escherichia coli E22] 163 YP_541301 91211315 6-phosphogluconate dehydrogenase, decarboxylating [Escherichia coli UTI89] 164 YP_013993 46907604 6-phosphogluconate dehydrogenase [Listeria monocytogenes str. 4b F2365] 165 YP_403788 82777439 gluconate-6-phosphate dehydrogenase [Shigella dysenteriae Sd197] 166 ZP_009 . . . 83588030 COG0362: 6-phosphogluconate dehydrogenase [Escherichia coli 101-1] 167 AAD50492 5739470 6-phosphogluconate dehydrogenase Gnd [Escherichia coli] 168 ZP_006 . . . 75177157 COG0362: 6-phosphogluconate dehydrogenase [Shigella boydii BS512] 169 AAA24209 146942 6-phosphogluconate dehydrogenase 170 AAG35221 11464522 6-phosphogluconate dehydrogenase [Escherichia coli] 171 ZP_007 . . . 75186836 COG0362: 6-phosphogluconate dehydrogenase [Escherichia coli E24377A] 172 YP_689521 110806001 gluconate-6-phosphate dehydrogenase [Shigella flexneri 5 str. 8401] 173 ZP_008 . . . 77959279 COG0362: 6-phosphogluconate dehydrogenase [Yersinia bercovieri ATCC 43970] 174 AAG35224 11464528 6-phosphogluconate dehydrogenase [Escherichia coli] 175 ZP_007 . . . 77636293 COG0362: 6-phosphogluconate dehydrogenase [Yersinia pestis Angola] 176 ZP_008 . . . 77962467 COG0362: 6-phosphogluconate dehydrogenase [Yersinia mollaretii ATCC 43969] 177 ZP_008 . . . 77978861 COG0362: 6-phosphogluconate dehydrogenase [Yersinia intermedia ATCC 29909] 178 AAG35218 11464516 6-phosphogluconate dehydrogenase [Escherichia coli] 179 BAA77736 4867927 6-phosphogluconate dehydrogenase [Escherichia coli] 180 AAA24490 147491 6-phosphogluconate dehydrogenase 181 YP_648367 108812600 6-phosphogluconate dehydrogenase, decarboxylating [Yersinia pestis Nepal516] 182 NP_669932 22126509 6-phosphogluconate dehydrogenase [Yersinia pestis KIM] 183 AAA23918 146232 6-phosphogluconate dehydrogenase (EC 1.1.1.44) 184 CAJ71042 91203389 strongly similar to 6-phosphogluconate dehydrogenase (decarboxylating) [Candidatus Kuenenia stuttgartiensis] 185 AAA24206 146936 6-phosphogluconate dehydrogenase 186 AAA24494 147499 6-phosphogluconate dehydrogenase 187 AAA24492 147495 6-phosphogluconate dehydrogenase 188 YP_099135 53713143 6-phosphogluconate dehydrogenase [Bacteroides fragilis YCH46] 189 AAA23925 146246 6-phosphogluconate dehydrogenase 190 YP_211552 60681408 6-phosphogluconate dehydrogenase [Bacteroides fragilis NCTC 9343] 191 AAV74381 56122509 Gnd [Escherichia coli] 192 NP_992794 45441255 6-phosphogluconate dehydrogenase [Yersinia pestis biovar Microtus str. 91001] 193 YP_650749 108806833 6-phosphogluconate dehydrogenase, decarboxylating [Yersinia pestis Antiqua] 194 NP_876643 33241702 6-phosphogluconate dehydrogenase [Chlamydophila pneumoniae TW-183] 195 NP_357929 15902379 6-phosphogluconate dehydrogenase [Streptococcus pneumoniae R6] 196 ZP_014 . . . 111658214 hypothetical protein SpneT_02000606 [Streptococcus pneumoniae TIGR4] 197 NP_782446 28211502 6-phosphogluconate dehydrogenase [Clostridium tetani E88] 198 AAG35223 11464526 6-phosphogluconate dehydrogenase [Escherichia coli] 199 AAA24493 147497 6-phosphogluconate dehydrogenase 200 ZP_007 . . . 75214880 COG0362: 6-phosphogluconate dehydrogenase [Escherichia coli E110019]

TABLE 47 Examples of Fructose 1,6 bisphosphatase polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_500111 50545147 hypothetical protein [Yarrowia lipolytica]. 2 AAP85294 32492043 fructose-1,6-bisphosphatase [Yarrowia lipolytica] 3 XP_460410 50423651 hypothetical protein DEHA0F01309g [Debaryomyces hansenii CBS767] 4 CAC69139 15528447 putative fructose-1,6-bisphosphatase [Pichia anomala] 5 XP_447425 50289987 hypothetical protein CAGL0I04048g [Candida glabrata CBS138] 6 NP_986140 45199111 AFR593Cp [Eremothecium gossypii] 7 XP_454003 50308005 F16P_KLULA [Kluyveromyces lactis] 8 EAT78213 111057093 predicted protein [Phaeosphaeria nodorum SN15] 9 XP_710934 68490480 fructose-1,6-bisphosphatase [Candida albicans SC5314] 10 AAT92835 51013083 YLR377C [Saccharomyces cerevisiae] 11 XP_364050 39954137 hypothetical protein MG08895.4 [Magnaporthe grisea 70-15] 12 NP_001 . . . 68006542 fructose-1,6-bisphosphatase [Schizosaccharomyces pombe 972h-] 13 NP_595083 19111875 hypothetical protein SPBC1198.14c [Schizosaccharomyces pombe 972h-] 14 XP_751698 70993702 fructose-1,6-bisphosphatase [Aspergillus fumigatus Af293] 15 XP_447010 50289161 unnamed protein product [Candida glabrata] 16 BAE58185 83768046 unnamed protein product [Aspergillus oryzae] 17 EAQ84924 88177456 conserved hypothetical protein [Chaetomium globosum CBS 148.51] 18 AAF34693 6980006 fructose 1,6-bisphosphatase [Candida albicans] 19 BAB12208 9955389 fructose-1,6-bisphosphatase [Aspergillus oryzae] 20 XP_663208 67538868 hypothetical protein AN5604.2 [Aspergillus nidulans FGSC A4] 21 XP_389456 46134281 conserved hypothetical protein [Gibberella zeae PH-1] 22 XP_960423 85097320 hypothetical protein [Neurospora crassa OR74A] 23 EAS36192 90306561 fructose-1,6-bisphosphatase [Coccidioides immitis RS] 24 XP_566475 58258125 fructose-bisphosphatase [Cryptococcus neoformans var. neoformans JEC21] 25 XP_758850 71015920 hypothetical protein UM02703.1 [Ustilago maydis 521] 26 P14766 119748 Fructose-1,6-bisphosphatase, cytosolic (D-fructose-1,6- bisphosphate 1-phosphohydrolase) (FBPase) 27 AAZ86538 73811203 fructose 1,6-bisphosphatase [Brassica rapa subsp. pekinensis] 28 Q9MA79 75312318 Fructose-1,6-bisphosphatase, cytosolic (D-fructose-1,6- bisphosphate 1-phosphohydrolase) (FBPase) 29 Q9SDL8 75313310 Fructose-1,6-bisphosphatase, cytosolic (D-fructose-1,6- bisphosphate 1-phosphohydrolase) (FBPase) 30 BAD81916 56785198 fructose-1,6-bisphosphatase [Oryza sativa (japonica cultivar- group)] 31 P46267 1169585 Fructose-1,6-bisphosphatase, cytosolic (D-fructose-1,6- bisphosphate 1-phosphohydrolase) (FBPase) 32 P46276 1169586 Fructose-1,6-bisphosphatase, cytosolic (D-fructose-1,6- bisphosphate 1-phosphohydrolase) (FBPase) (CY-F1) 33 Q8RW99 75303233 Fructose-1,6-bisphosphatase, cytosolic (D-fructose-1,6- bisphosphate 1-phosphohydrolase) (FBPase) 34 Q43139 76363515 Fructose-1,6-bisphosphatase, cytosolic (D-fructose-1,6- bisphosphate 1-phosphohydrolase) (FBPase) 35 CAB46084 5305145 fructose-1,6-bisphosphatase [Pisum sativum] 36 BAF06820 113534437 Os01g0866400 [Oryza sativa (japonica cultivar-group)] 37 Q42649 76789650 Fructose-1,6-bisphosphatase, cytosolic (D-fructose-1,6- bisphosphate 1-phosphohydrolase) (FBPase) 38 Q9XF47 75315047 Fructose-1,6-bisphosphatase, cytosolic (D-fructose-1,6- bisphosphate 1-phosphohydrolase) (FBPase) 39 CAA61409 895909 fructose-1,6-bisphosphatase [Saccharum hybrid cultivar H65- 7052] 40 1906373A 444324 cytosolic fructose bisphosphatase 41 EAR87347 89289359 fructose-1,6-bisphosphatase family protein [Tetrahymena thermophila SB210] 42 NP_915641 34908588 fructose-1,6-bisphosphatase [Oryza sativa (japonica cultivar- group)] 43 XP_475314 50931573 putative fructose-1,6-bisphosphatase [Oryza sativa (japonica cultivar-group)] 44 NP_491004 17508131 K07A3.1 [Caenorhabditis elegans] 45 XP_782411 72013871 PREDICTED: similar to fructose-1,6-bisphosphatase 1, like [Strongylocentrotus purpuratus] 46 CAE60538 39595500 Hypothetical protein CBG04165 [Caenorhabditis briggsae] 47 NP_610001 19921562 fructose-1,6-bisphosphatase CG31692-PA, isoform A [Drosophila melanogaster] 48 NP_724223 45550998 fructose-1,6-bisphosphatase CG31692-PB, isoform B [Drosophila melanogaster] 49 XP_319937 58393257 ENSANGP00000016841 [Anopheles gambiae str. PEST] 50 AAN31471 23394363 fructose-1 6-biphosphatase [Phytophthora infestans] 51 XP_425040 50762393 PREDICTED: similar to fructose 1,6-bisphosphatase [Gallus gallus] 52 EAT47721 108883496 fructose-1,6-bisphosphatase [Aedes aegypti] 53 EAT47722 108883497 fructose-1,6-bisphosphatase [Aedes aegypti] 54 AAH53784 32450356 Fbp-prov protein [Xenopus laevis] 55 YP_001659 45657573 fructose-1,6-bisphosphatase [Leptospira interrogans serovar Copenhageni str. Fiocruz L1-130] 56 NP_989145 45361017 Fructose-1,6-bisphosphatase [Xenopus tropicalis] 57 1YZ0B 62738852 Chain B, R-State Amp Complex Reveals Initial Steps Of The Quaternary Transition Of Fructose-1,6-Bisphosphatase 58 1Q9DB 39654661 Chain B, Fructose-1,6-Bisphosphatase Complexed With A New Allosteric Site Inhibitor (I-State) 59 NP_999144 47522784 fructose 1,6-bisphosphatase [Sus scrofa] 60 EAL32807 54644065 GA16400-PA [Drosophila pseudoobscura] 61 XP_848306 73946392 PREDICTED: similar to fructose-1,6-bisphosphatase 1 isoform 3 [Canis familiaris] 62 1FJ9B 11514534 Chain B, Fructose-1,6-Bisphosphatase (Mutant Y57w) ProductsZNAMP Complex (T-State) 63 1LEVF 24987566 Chain F, Porcine Kidney Fructose-1,6-Bisphosphatase Complexed With An Amp-Site Inhibitor 64 1BK4A 6729708 Chain A, Crystal Structure Of Rabbit Liver Fructose-1,6- Bisphosphatase At 2.3 Angstrom Resolution 65 Q3SZB7 110808224 Fructose-1,6-bisphosphatase 1 (D-fructose-1,6-bisphosphate 1-phosphohydrolase 1) (FBPase 1) 66 AAA41131 310111 fructose-biphosphatase 67 P46275 2506391 Fructose-1,6-bisphosphatase, chloroplast precursor (D- fructose-1,6-bisphosphate 1-phosphohydrolase) (FBPase) 68 2F3HB 99032142 Chain B, Mechanism Of Displacement Of A Catalytically Essential Loop From The Active Site Of Fructose-1,6- Bisphosphatase 69 NP_036690 51036635 fructose-1,6-biphosphatase 1 [Rattus norvegicus] 70 AAC25597 3288991 fructose-1,6-bisphosphatase [Sus scrofa] 71 XP_967802 91077758 PREDICTED: similar to CG31692-PA, isoform A [Tribolium castaneum] 72 AAK59929 14318171 fructose-1,6-bisphosphatase [Pisum sativum] 73 1DCUD 6730314 Chain D, Redox Signaling In The Chloroplast: Structure Of Oxidized Pea Fructose-1,6-Bisphosphate Phosphatase 74 1DBZD 6730306 Chain D, C153s Mutant Of Pea Fructose-1,6-Bisphosphatase 75 CAB39759 4539148 fructose-1,6-bisphosphatase [Pisum sativum] 76 P09199 20141075 Fructose-1,6-bisphosphatase 1 (D-fructose-1,6-bisphosphate 1-phosphohydrolase 1) (FBPase 1) 77 CAA48719 20717 fructose-bisphosphatase [Pisum sativum] 78 1FPIB 1633402 Chain B, Fructose-1,6-Bisphosphatase (D-Fructose-1,6- Bisphosphate 1-Phosphohydrolase) Complexed With Amp, 2,5-Anhydro-D-Glucitol-1,6-Bisphosphate And Potassium Ions (100 Mm) 79 AAB30523 546354 fructose-1,6-biphosphatase, FBPase {EC 3.1.3.11} [Pisum sativum = peas, Lincoln, Peptide Chloroplast, 357 aa] 80 1KZ8F 24987484 Chain F, Crystal Structure Of Porcine Fructose-1,6- Bisphosphatase Complexed With A Novel Allosteric-Site Inhibitor 81 AAW25416 56754457 SJCHGC06706 protein [Schistosoma japonicum] 82 1FRPB 809402 Chain B, Fructose-1,6-Bisphosphatase (D-Fructose-1,6- Bisphosphate 1-Phosphohydrolase) (E.C.3.1.3.11) Complexed With Fructose-2,6-Bisphosphate, Adenosine Monophosphate (Amp), And Zinc 83 XP_425039 50762391 PREDICTED: similar to fructose-1,6-bisphosphatase [Gallus gallus] 84 NP_998297 47085885 fructose-1,6-bisphosphatase 1 [Danio rerio] 85 1RDZB 1942591 Chain B, T-State Structure Of The Arg 243 To Ala Mutant Of Pig Kidney Fructose 1,6-Bisphosphatase Expressed In E. Coli 86 1FSAB 2554913 Chain B, The T-State Structure Of Lys 42 To Ala Mutant Of The Pig Kidney Fructose 1,6-Bisphosphatase Expressed In E. Coli 87 BAE28940 74146344 unnamed protein product [Mus musculus] 88 CAG05216 47228396 unnamed protein product [Tetraodon nigroviridis] 89 NP_956236 41053949 fructose-1,6-bisphosphatase 1, like [Danio rerio] 90 Q07204 585118 Fructose-1,6-bisphosphatase, chloroplast precursor (D- fructose-1,6-bisphosphate 1-phosphohydrolase) (FBPase) 91 BAF11578 113548135 Os03g0267300 [Oryza sativa (japonica cultivar-group)] 92 2FIXL 90109451 Chain L, Structure Of Human Liver Fbpase Complexed With Potent Benzoxazole Allosteric Inhibitiors 93 2FHYL 90109424 Chain L, Structure Of Human Liver Fpbase Complexed With A Novel Benzoxazole As Allosteric Inhibitor 94 XP_001 . . . 109112377 PREDICTED: fructose-1,6-bisphosphatase 1 isoform 2 [Macaca mulatta] 95 1FTAD 1311149 Chain D, Fructose-1,6-Bisphosphatase(D-Fructose-1,6- Bisphosphate, 1-Phosphohydrolase) (E.C.3.1.3.11) Complexed With The Allosteric Inhibitor Amp 96 CAH72692 55662224 fructose-1,6-bisphosphatase 1 [Homo sapiens] 97 CAG08190 47221528 unnamed protein product [Tetraodon nigroviridis] 98 XP_533504 73946534 PREDICTED: similar to Fructose-1,6-bisphosphatase isozyme 2 (D-fructose-1,6-bisphosphate 1-phosphohydrolase) (FBPase) [Canis familiaris] 99 AAC25774 3293553 fructose-1,6-bisphosphatase [Homo sapiens] 100 AAA35817 182311 fructose-1,6-bisphosphatase 101 AAD12243 4218951 fructose-1,6-bisphosphatase precursor [Brassica napus] 102 NP_001 . . . 114051459 hypothetical protein LOC514066 [Bos taurus] 103 P09195 119745 Fructose-1,6-bisphosphatase, chloroplast precursor (D- fructose-1,6-bisphosphate 1-phosphohydrolase) (FBPase) 104 NP_001 . . . 57524507 hypothetical protein LOC445505 [Danio rerio] 105 NP_001 . . . 114051287 fructose-1,6-bisphosphatase [Bombyx mori] 106 NP_001 . . . 73853862 hypothetical protein LOC613108 [Xenopus tropicalis] 107 AAD25541 4585576 fructose-1,6-bisphosphatase precursor [Solanum tuberosum] 108 Q9N0J6 75067927 Fructose-1,6-bisphosphatase isozyme 2 (D-fructose-1,6- bisphosphate 1-phosphohydrolase 2) (FBPase 2) 109 AAN31884 23397203 putative fructose-bisphosphatase precursor [Arabidopsis thaliana] 110 AAH81229 51703601 Unknown (protein for MGC: 85456) [Xenopus laevis] 111 P22418 3915687 Fructose-1,6-bisphosphatase, chloroplast precursor (D- fructose-1,6-bisphosphate 1-phosphohydrolase) (FBPase) 112 CAA41154 11242 fructose-bisphosphatase [Arabidopsis thaliana] 113 XP_001 . . . 109112375 PREDICTED: fructose-1,6-bisphosphatase 1 isoform 1 [Macaca mulatta] 114 P70695 76363514 Fructose-1,6-bisphosphatase isozyme 2 (D-fructose-1,6- bisphosphate 1-phosphohydrolase 2) (FBPase 2) (RAE-30) 115 CAG08189 47221527 unnamed protein product [Tetraodon nigroviridis] 116 AAI13633 109731177 Fructose-1,6-bisphosphatase 2 [Homo sapiens] 117 1SPID 999624 Chain D, Fructose-1,6-Bisphosphatase (D-Fructose-1,6- Bisphosphate 1-Phosphohydrolase) (E.C.3.1.3.11) 118 AAF72973 8118281 fructose-1,6-bisphosphatase [Zaocys dhumnades] 119 AAH12720 15215255 Fbp2 protein [Mus musculus] 120 O00757 76789651 Fructose-1,6-bisphosphatase isozyme 2 (D-fructose-1,6- bisphosphate 1-phosphohydrolase 2) (FBPase 2) 121 XP_827196 71745132 fructose-1,6-bisphosphate cytosolic [Trypanosoma brucei TREU927] 122 Q9Z1N1 76364186 Fructose-1,6-bisphosphatase isozyme 2 (D-fructose-1,6- bisphosphate 1-phosphohydrolase 2) (FBPase 2) 123 BAE40306 74212301 unnamed protein product [Mus musculus] 124 CAC70747 15718373 fructose-1,6-bisphosphatase [Trypanosoma brucei] 125 NP_032020 6679761 fructose bisphosphatase 2 [Mus musculus] 126 AAP79192 32307574 fructose-1,6 bisphosphatase [Bigelowiella natans] 127 ZP_010 . . . 86130901 fructose-1,6-bisphosphatase [Cellulophaga sp. MED134] 128 YP_422024 83311760 Fructose-1,6-bisphosphatase [Magnetospirillum magneticum AMB-1] 129 YP_679484 110639275 fructose-1,6-bisphosphatase [Cytophaga hutchinsonii ATCC 33406] 130 ZP_000 . . . 23014308 COG0158: Fructose-1,6-bisphosphatase [Magnetospirillum magnetotacticum MS-1] 131 XP_888627 76363832 Leishmania major strain Friedlin cytosolic fructose-1,6- bisphosphatase [Leishmania major strain Friedlin] 132 ZP_009 . . . 83855881 fructose-1,6-bisphosphatase [Croceibacter atlanticus HTCC2559] 133 YP_437516 83649081 Fructose-1,6-bisphosphatase [Hahella chejuensis KCTC 2396] 134 ZP_011 . . . 88711300 fructose-1,6-bisphosphatase [Flavobacteriales bacterium HTCC2170] 135 CAC82800 16519317 fructose 1,6-bisphosphatase [Galdieria sulphuraria] 136 ZP_012 . . . 91215105 fructose-1,6-bisphosphatase [Psychroflexus torquis ATCC 700755] 137 ZP_012 . . . 90591576 Inositol phosphatase/fructose-1,6-bisphosphatase [Flavobacterium johnsoniae UW101] 138 XP_793452 72046390 PREDICTED: similar to fructose-1,6-bisphosphatase 1, like [Strongylocentrotus purpuratus] 139 CAJ72223 91202584 strongly similar to fructose-1,6-bisphosphatase [Candidatus Kuenenia stuttgartiensis] 140 ZP_011 . . . 88804197 fructose-1,6-bisphosphatase [Robiginitalea biformata HTCC2501] 141 YP_445598 83815956 fructose-1,6-bisphosphatase [Salinibacter ruber DSM 13855] 142 ZP_007 . . . 75815977 COG0158: Fructose-1,6-bisphosphatase [Vibrio cholerae V52] 143 ZP_011 . . . 88802508 fructose-1,6-bisphosphatase [Polaribacter irgensii 23-P] 144 ZP_007 . . . 75827798 COG0158: Fructose-1,6-bisphosphatase [Vibrio cholerae O395] 145 ZP_011 . . . 88797163 Fructose-1,6-bisphosphatase [Reinekea sp. MED297] 146 ZP_005 . . . 68553442 Inositol phosphatase/fructose-1,6-bisphosphatase [Prosthecochloris aestuarii DSM 271] 147 ZP_007 . . . 75830525 COG0158: Fructose-1,6-bisphosphatase [Vibrio cholerae MO10] 148 Q42796 2494415 Fructose-1,6-bisphosphatase, chloroplast precursor (D- fructose-1,6-bisphosphate 1-phosphohydrolase) (FBPase) 149 ZP_007 . . . 75822783 COG0158: Fructose-1,6-bisphosphatase [Vibrio cholerae RC385] 150 NP_933227 37678618 fructose-1,6-bisphosphatase [Vibrio vulnificus YJ016] 151 XP_812636 71423971 fructose-1,6-bisphosphatase, cytosolic [Trypanosoma cruzi strain CL Brener] 152 ZP_006 . . . 71480926 Inositol phosphatase/fructose-1,6-bisphosphatase [Prosthecochloris vibrioformis DSM 265] 153 ZP_007 . . . 75854570 COG0158: Fructose-1,6-bisphosphatase [Vibrio sp. Ex25] 154 NP_931714 37528369 fructose-1,6-bisphosphatase [Photorhabdus luminescens subsp. laumondii TTO1] 155 XP_805839 71406635 fructose-1,6-bisphosphatase, cytosolic [Trypanosoma cruzi strain CL Brener] 156 YP_374392 78186349 fructose-1,6-bisphosphatase [Pelodictyon luteolum DSM 273] 157 NP_902097 34497882 fructose-1,6-bisphosphatase [Chromobacterium violaceum ATCC 12472] 158 XP_646323 66825937 D-fructose-1,6-bisphosphate 1-phosphohydrolase [Dictyostelium discoideum AX4] 159 ZP_010 . . . 86147594 fructose-1,6-bisphosphatase [Vibrio sp. MED222] 160 NP_796691 28897086 fructose-1,6-bisphosphatase [Vibrio parahaemolyticus RIMD 2210633] 161 YP_052014 50122847 fructose-1,6-bisphosphatase [Erwinia carotovora subsp. atroseptica SCRI1043] 162 ZP_007 . . . 77634762 COG0158: Fructose-1,6-bisphosphatase [Yersinia pestis Angola] 163 ZP_012 . . . 90406722 fructose-1,6-bisphosphatase [Psychromonas sp. CNPT3] 164 ZP_006 . . . 71549000 Inositol phosphatase/fructose-1,6-bisphosphatase [Nitrosomonas eutropha C71] 165 ZP_013 . . . 106882698 Inositol phosphatase/fructose-1,6-bisphosphatase [Psychromonas ingrahamii 37] 166 ZP_008 . . . 77973994 COG0158: Fructose-1,6-bisphosphatase [Yersinia frederiksenii ATCC 33641] 167 ZP_012 . . . 90411963 fructose-1,6-bisphosphatase [Photobacterium profundum 3TCK] 168 ZP_005 . . . 67938188 Inositol phosphatase/fructose-1,6-bisphosphatase [Chlorobium phaeobacteroides BS1] 169 NP_991956 45440417 fructose-1,6-bisphosphatase [Yersinia pestis biovar Microtus str. 91001] 170 NP_840606 30248536 fructose-1,6-bisphosphatase [Nitrosomonas europaea ATCC 19718] 171 YP_649191 108813424 fructose-1,6-bisphosphatase [Yersinia pestis Nepal516] 172 ZP_010 . . . 86134593 fructose-1,6-bisphosphatase [Tenacibaculum sp. MED152] 173 ZP_012 . . . 94268582 Inositol phosphatase/fructose-1,6-bisphosphatase [delta proteobacterium MLMS-1] 174 ZP_012 . . . 94264279 Inositol phosphatase/fructose-1,6-bisphosphatase [delta proteobacterium MLMS-1] 175 ZP_005 . . . 67919158 Inositol phosphatase/fructose-1,6-bisphosphatase [Chlorobium limicola DSM 245] 176 NP_661262 21673197 fructose-1,6-bisphosphatase [Chlorobium tepidum TLS] 177 YP_128618 54307598 fructose-1,6-bisphosphatase [Photobacterium profundum SS9] 178 YP_378918 78188580 fructose-1,6-bisphosphatase [Chlorobium chlorochromatii CaD3] 179 ZP_005 . . . 68551142 Inositol phosphatase/fructose-1,6-bisphosphatase [Pelodictyon phaeoclathratiforme BU-1] 180 YP_445968 83814754 fructose-1,6-bisphosphatase [Salinibacter ruber DSM 13855] 181 ZP_012 . . . 90580816 fructose-1,6-bisphosphatase [Vibrio angustum S14] 182 ZP_011 . . . 89075542 fructose-1,6-bisphosphatase [Photobacterium sp. SKA34] 183 YP_342093 77163568 fructose-1,6-bisphosphatase [Nitrosococcus oceani ATCC 19707] 184 ZP_009 . . . 84387793 fructose-1,6-bisphosphatase [Vibrio splendidus 12B01] 185 ZP_010 . . . 86141292 fructose-1,6-bisphosphatase [Flavobacterium sp. MED217] 186 ZP_005 . . . 67937321 Inositol phosphatase/fructose-1,6-bisphosphatase [Chlorobium phaeobacteroides DSM 266] 187 ZP_012 . . . 91228829 fructose-1,6-bisphosphatase [Vibrio alginolyticus 12G01] 188 ZP_008 . . . 77961846 COG0158: Fructose-1,6-bisphosphatase [Yersinia mollaretii ATCC 43969] 189 ZP_008 . . . 77977605 COG0158: Fructose-1,6-bisphosphatase [Yersinia intermedia ATCC 29909] 190 YP_203647 59710871 fructose-1,6-bisphosphatase [Vibrio fischeri ES114] 191 ZP_013 . . . 110596824 Inositol phosphatase/fructose-1,6-bisphosphatase [Chlorobium ferrooxidans DSM 13031] 192 YP_454036 85058334 fructose-1,6-bisphosphatase [Sodalis glossinidius str. ‘morsitans’] 193 ZP_001 . . . 32035025 COG0158: Fructose-1,6-bisphosphatase [Actinobacillus pleuropneumoniae serovar 1 str. 4074] 194 YP_011058 46580250 fructose-1,6-bisphosphatase [Desulfovibrio vulgaris subsp. vulgaris str. Hildenborough] 195 YP_219276 62182859 fructose-1,6-bisphosphatase [Salmonella enterica subsp. enterica serovar Choleraesuis str. SC-B67] 196 YP_560077 91784871 Inositolphosphatase/fructose-1,6-bisphosphatase [Burkholderia xenovorans LB400] 197 YP_316335 74318595 fructose-1,6-bisphosphatase [Thiobacillus denitrificans ATCC 25259] 198 YP_388561 78357112 fructose-1,6-bisphosphatase [Desulfovibrio desulfuricans G20] 199 NP_313236 15834463 fructose-1,6-bisphosphatase [Escherichia coli O157:H7 str. Sakai] 200 YP_313128 74314709 fructose-bisphosphatase [Shigella sonnei Ss046] 201 ZP_011 . . . 88801533 fructose-1,6-bisphosphatase [Polaribacter irgensii 23-P]

TABLE 53 Examples of squalene epoxidase polypeptides Row ACCESSION GI PROTEIN DESCRIPTION 1 XP_503994 50553168 hypothetical protein [Yarrowia lipolytica]. 2 EAS30670 90301039 hypothetical protein CIMG_06149 [Coccidioides immitis RS] 3 XP_753858 70998268 squalene monooxygenase Erg1 [Aspergillus fumigatus Af293] 4 XP_711894 68488538 squalene epoxidase [Candida albicans SC5314] 5 ABD48706 88697124 squalene epoxidase [Emericella nidulans] 6 XP_681020 67901528 hypothetical protein AN7751.2 [Aspergillus nidulans FGSC A4] 7 AAQ18215 33415411 squalene epoxidase [Trichophyton rubrum] 8 BAE62359 83772229 unnamed protein product [Aspergillus oryzae] 9 AAQ18216 61807216 squalene epoxidase [Trichophyton rubrum] 10 AAZ08563 70797544 squalene epoxidase [Trichophyton rubrum] 11 CAJ18281 90185988 squalene epoxidase [Hypocrea lixii] 12 XP_462160 50427105 hypothetical protein DEHA0G15202g [Debaryomyces hansenii CBS767] 13 NP_011691 6321614 Squalene epoxidase, catalyzes the epoxidation of squalene to 2,3- oxidosqualene; plays an essential role in the ergosterol-biosynthesis pathway and is the specific target of the antifungal drug terbinafine; Erg1p [Saccharomyces cerevisiae] 14 AAA34592 171471 squalene epoxidase 15 O13306 51704232 Squalene monooxygenase (Squalene epoxidase) (SE) 16 XP_455763 50311477 unnamed protein product [Kluyveromyces lactis] 17 XP_386391 46123675 hypothetical protein FG06215.1 [Gibberella zeae PH-1] 18 EAQ88139 88180671 hypothetical protein CHGG_04758 [Chaetomium globosum CBS 148.51] 19 XP_961806 85104779 hypothetical protein [Neurospora crassa OR74A] 20 XP_758545 71012984 hypothetical protein UM02398.1 [Ustilago maydis 521] 21 XP_369325 39975869 hypothetical protein MG06139.4 [Magnaporthe grisea 70-15] 22 XP_570092 58265872 squalene monooxygenase [Cryptococcus neoformans var. neoformans JEC21] 23 Q9C1W3 51701448 Probable squalene monooxygenase (Squalene epoxidase) (SE) 24 EAT79729 111058609 hypothetical protein SNOG_12929 [Phaeosphaeria nodorum SN15] 25 CAG38355 108743007 squalene epoxidase [Aspergillus niger] 26 BAE73094 84579321 hypothetical protein [Macaca fascicularis] 27 EAU31524 114189824 hypothetical protein ATEG_08351 [Aspergillus terreus NIH2624] 28 NP_003120 62865635 squalene monooxygenase [Homo sapiens] 29 CAI46076 57997512 hypothetical protein [Homo sapiens] 30 BAE37374 74200967 unnamed protein product [Mus musculus] 31 XP_850640 73974448 PREDICTED: similar to squalene monooxygenase isoform 1 [Canis familiaris] 32 Q75F69 51701415 Squalene monooxygenase (Squalene epoxidase) (SE) 33 AAS60234 45388098 squalene epoxidase 1 [Aspergillus fumigatus] 34 Q14534 12644414 Squalene monooxygenase (Squalene epoxidase) (SE) 35 XP_001 . . . 109087424 PREDICTED: similar to squalene monooxygenase [Macaca mulatta] 36 AAH97330 66911977 Sqle protein [Rattus norvegicus] 37 EAU30782 114189082 hypothetical protein ATEG_08650 [Aspergillus terreus NIH2624] 38 CAF98057 47218525 unnamed protein product [Tetraodon nigroviridis] 39 XP_861944 73974450 PREDICTED: similar to squalene monooxygenase isoform 2 [Canis familiaris] 40 ABF94794 108706999 Squalene monooxygenase, putative, expressed [Oryza sativa (japonica cultivar-group)] 41 BAF11377 113547934 Os03g0231800 [Oryza sativa (japonica cultivar-group)] 42 XP_470614 50920507 Putative Squalene monooxygenase [Oryza sativa (japonica cultivar- group)] 43 ABF94791 108706996 Squalene monooxygenase, putative, expressed [Oryza sativa (japonica cultivar-group)] 44 AAM61384 21537043 squalene epoxidase-like protein [Arabidopsis thaliana] 45 BAE98940 110742002 hypothetical protein [Arabidopsis thaliana] 46 ABE84801 92877833 Helix-turn-helix, AraC type; NAD-binding site; Fumarate lyase [Medicago truncatula] 47 BAE36598 74183459 unnamed protein product [Mus musculus] 48 XP_693698 68403773 PREDICTED: similar to squalene monooxygenase [Danio rerio] 49 CAB80441 7270759 squalene epoxidase-like protein [Arabidopsis thaliana] 50 O48651 6685403 Squalene monooxygenase (Squalene epoxidase) (SE) 51 AAY22200 62945915 squalene monooxygenase [Datura innoxia] 52 BAD15330 46359651 squalene epoxidase [Panax ginseng] 53 CAD23248 27475610 squalene monooxygenase 2 [Medicago truncatula] 54 ABC94943 86371770 squalene epoxidase [Medicago sativa] 55 CAD23249 27475612 squalene monooxygenase 1 [Medicago truncatula] 56 AAN15558 23198062 squalene monooxygenase, putative [Arabidopsis thaliana] 57 NP_179868 15227757 oxidoreductase [Arabidopsis thaliana] 58 AAG50645 12321049 squalene monooxygenase, putative [Arabidopsis thaliana] 59 XP_629022 66800193 hypothetical protein DDBDRAFT_0192021 [Dictyostelium discoideum AX4] 60 CAK11466 94733753 novel protein similar to vertebrate squalene epoxidase (SQLE) [Danio rerio] 61 AAQ13595 33337947 putative squalene epoxidase [Lycopersicon esculentum] 62 ABF94792 108706997 Squalene monooxygenase, putative, expressed [Oryza sativa (japonica cultivar-group)] 63 XP_604908 76660360 PREDICTED: similar to squalene monooxygenase [Bos taurus] 64 BAA11209 1483186 squalene epoxidase [Homo sapiens] 65 NP_197803 15237902 SQP1 [Arabidopsis thaliana] 66 NP_001 . . . 79328611 SQP1 [Arabidopsis thaliana] 67 CAA06772 3123331 squalene epoxidase homologue [Arabidopsis thaliana] 68 O65726 6685410 Squalene monooxygenase 1,2 (Squalene epoxidase 1,2) (SE 1,2) 69 O65727 6685411 Squalene monooxygenase 1,1 (Squalene epoxidase 1,1) (SE 1,1) 70 NP_197804 15237903 oxidoreductase [Arabidopsis thaliana] 71 CAJ03182 68125205 squalene monooxygenase-like protein [Leishmania major] 72 NP_197802 15237900 SQP2; oxidoreductase [Arabidopsis thaliana] 73 XP_828409 71754989 squalene monooxygenase [Trypanosoma brucei TREU927] 74 XP_808343 71412304 squalene monooxygenase [Trypanosoma cruzi strain CL Brener] 75 XP_789059 72038634 PREDICTED: similar to squalene monooxygenase [Strongylocentrotus purpuratus] 76 XP_796297 72133073 PREDICTED: similar to squalene monooxygenase [Strongylocentrotus purpuratus] 77 AAB69189 2352528 squalene epoxidase [Candida glabrata] 78 XP_813434 71649420 squalene monooxygenase [Trypanosoma cruzi strain CL Brener] 79 XP_796261 72133071 PREDICTED: similar to squalene monooxygenase [Strongylocentrotus purpuratus] 80 XP_793783 72114550 PREDICTED: similar to squalene monooxygenase, partial [Strongylocentrotus purpuratus] 81 XP_789028 72038632 PREDICTED: similar to squalene xygenase [Strongylocentrotus purpuratus] 82 AAT97087 51105054 squalene epoxidase-like protein [Lymnaea stagnalis] 83 XP_804343 71402993 squalene monooxygenase [Trypanosoma cruzi strain CL Brener] 84 YP_115265 53803024 monooxygenase family protein [Methylococcus capsulatus str. Bath] 85 AAM91177 22136198 unknown protein [Arabidopsis thaliana] 86 XP_799134 72026766 PREDICTED: similar to Squalene monooxygenase (Squalene epoxidase) (SE), partial [Strongylocentrotus purpuratus]

TABLE 54 Examples of ERG1 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAA34592 171471 squalene epoxidase 2 O13306 51704232 Squalene monooxygenase (Squalene epoxidase) (SE) 3 XP_455763 50311477 unnamed protein product [Kluyveromyces lactis] 4 Q75F69 51701415 Squalene monooxygenase (Squalene epoxidase) (SE) 5 XP_462160 50427105 hypothetical protein DEHA0G15202g [Debaryomyces hansenii CBS767] 6 XP_711894 68488538 squalene epoxidase [Candida albicans SC5314] 7 XP_503994 50553168 hypothetical protein [Yarrowia lipolytica] 8 AAB69189 2352528 squalene epoxidase [Candida glabrata] 9 AAQ18215 33415411 squalene epoxidase [Trichophyton rubrum] 10 AAQ18216 61807216 squalene epoxidase [Trichophyton rubrum] 11 AAZ08563 70797544 squalene epoxidase [Trichophyton rubrum] 12 EAS30670 90301039 hypothetical protein CIMG_06149 [Coccidioides immitis RS] 13 Q9C1W3 51701448 Probable squalene monooxygenase (Squalene epoxidase) (SE) 14 XP_758545 71012984 hypothetical protein UM02398.1 [Ustilago maydis 521] 15 XP_681020 67901528 hypothetical protein AN7751.2 [Aspergillus nidulans FGSC A4] 16 ABD48706 88697124 squalene epoxidase [Emericella nidulans] 17 BAE62359 83772229 unnamed protein product [Aspergillus oryzae] 18 XP_753858 70998268 squalene monooxygenase Erg1 [Aspergillus fumigatus Af293] 19 XP_961806 85104779 hypothetical protein [Neurospora crassa OR74A] 20 XP_386391 46123675 hypothetical protein FG06215.1 [Gibberella zeae PH-1] 21 CAJ18281 90185988 squalene epoxidase [Hypocrea lixii] 22 BAE73094 84579321 hypothetical protein [Macaca fascicularis] 23 BAE37374 74200967 unnamed protein product [Mus musculus] 24 XP_570092 58265872 squalene monooxygenase [Cryptococcus neoformans var.neoformans JEC21] 25 NP_003120 62865635 squalene monooxygenase [Homo sapiens] 26 EAT79729 111058609 hypothetical protein SNOG_12929 [Phaeosphaeria nodorum SN15] 27 AAH97330 66911977 Sqle protein [Rattus norvegicus] 28 CAI46076 57997512 hypothetical protein [Homo sapiens] 29 CAF98057 47218525 unnamed protein product [Tetraodon nigroviridis] 30 XP_850640 73974448 PREDICTED: similar to squalene monooxygenase isoform 1 [Canis familiaris] 31 XP_369325 39975869 hypothetical protein MG06139.4 [Magnaporthe grisea 70-15] 32 Q14534 12644414 Squalene monooxygenase (Squalene epoxidase) (SE) 33 XP_001 . . . 109087424 PREDICTED: similar to squalene monooxygenase [Macaca mulatta] 34 EAQ88139 88180671 hypothetical protein CHGG_04758 [Chaetomium globosum CBS 148.51] 35 XP_693698 68403773 PREDICTED: similar to squalene monooxygenase [Danio rerio] 36 CAG38355 108743007 squalene epoxidase [Aspergillus niger] 37 XP_861944 73974450 PREDICTED: similar to squalene monooxygenase isoform 2 [Canis familiaris] 38 XP_519950 55631226 PREDICTED: similar to squalene epoxidase [Pan troglodytes] 39 AAS60234 45388098 squalene epoxidase 1 [Aspergillus fumigatus] 40 XP_629022 66800193 hypothetical protein DDBDRAFT_0192021 [Dictyostelium discoideum AX4] 41 CAK11466 94733753 novel protein similar to vertebrate squalene epoxidase (SQLE) [Danio rerio] 42 BAE98940 110742002 hypothetical protein [Arabidopsis thaliana] 43 AAM61384 21537043 squalene epoxidase-like protein [Arabidopsis thaliana] 44 ABC94943 86371770 squalene epoxidase [Medicago sativa] 45 CAB80441 7270759 squalene epoxidase-like protein [Arabidopsis thaliana] 46 CAD23248 27475610 squalene monooxygenase 2 [Medicago truncatula] 47 BAE36598 74183459 unnamed protein product [Mus musculus] 48 AAQ13595 33337947 putative squalene epoxidase [Lycopersicon esculentum] 49 AAY22200 62945915 squalene monooxygenase [Datura innoxia] 50 ABF94791 108706996 Squalene monooxygenase, putative, expressed [Oryza sativa (japonica cultivar-group)] 51 AAN15558 23198062 squalene monooxygenase, putative [Arabidopsis thaliana] 52 ABF94794 108706999 Squalene monooxygenase, putative, expressed [Oryza sativa (japonica cultivar-group)] 53 XP_470614 50920507 Putative Squalene monooxygenase [Oryza sativa (japonica cultivar-group)] 54 BAF11377 113547934 Os03g0231800 [Oryza sativa (japonica cultivar-group)] 55 CAD23249 27475612 squalene monooxygenase 1 [Medicago truncatula] 56 AAG50645 12321049 squalene monooxygenase, putative [Arabidopsis thaliana] 57 NP_179868 15227757 oxidoreductase [Arabidopsis thaliana] 58 ABE84801 92877833 Helix-turn-helix, AraC type; NAD-binding site; Fumarate lyase [Medicago truncatula] 59 ABF94792 108706997 Squalene monooxygenase, putative, expressed [Oryza sativa (japonica cultivar-group)] 60 BAA11209 1483186 squalene epoxidase [Homo sapiens] 61 XP_604908 76660360 PREDICTED: similar to squalene monooxygenase [Bos taurus] 62 XP_828409 71754989 squalene monooxygenase [Trypanosoma brucei TREU927] 63 XP_808343 71412304 squalene monooxygenase [Trypanosoma cruzi strain CL Brener] 64 CAJ03182 68125205 squalene monooxygenase-like protein [Leishmania major] 65 O65726 6685410 Squalene monooxygenase 1,2 (Squalene epoxidase 1,2) (SE 1,2) 66 NP_197804 15237903 oxidoreductase [Arabidopsis thaliana] 67 O65727 6685411 Squalene monooxygenase 1,1 (Squalene epoxidase 1,1) (SE 1,1) 68 CAA06772 3123331 squalene epoxidase homologue [Arabidopsis thaliana] 69 NP_001 . . . 79328611 SQP1 [Arabidopsis thaliana] 70 NP_197803 15237902 SQP1 [Arabidopsis thaliana] 71 NP_197802 15237900 SQP2; oxidoreductase [Arabidopsis thaliana]

TABLE 55 Examples of ERG7 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAT93062 51013537 YHR072W [Saccharomyces cerevisiae] 2 AAA16975 465105 lanosterol synthase 3 XP_448182 50291499 unnamed protein product [Candida glabrata] 4 XP_451982 50304065 unnamed protein product [Kluyveromyces lactis] 5 NP_987017 45201447 AGR351Wp [Eremothecium gossypii] 6 XP_457937 50418833 hypothetical protein DEHA0C06501g [Debaryomyces hansenii CBS767] 7 XP_722471 68467112 2,3-oxidosqualene-lanosterol cyclase [Candida albicans SC5314] 8 XP_722612 68466833 2,3-oxidosqualene-lanosterol cyclase [Candida albicans SC5314] 9 1903190A 383419 oxidosqualene cyclase 10 XP_504990 50555163 hypothetical protein [Yarrowia lipolytica] 11 AAB08472 1549343 inactive oxidosqualene-lansoterol cyclase [Saccharomyces cerevisiae] 12 Q96WJ0 30913110 Lanosterol synthase (Oxidosqualene--lanosterol cyclase) (2,3- epoxysqualene--lanosterol cyclase) (OSC) 13 XP_747936 70984860 oxidosqualene:lanosterol cyclase [Aspergillus fumigatus Af293] 14 BAE61626 83771494 unnamed protein product [Aspergillus oryzae] 15 NP_593702 63054562 lanosterol synthase activity [Schizosaccharomyces pombe 972h-] 16 XP_758387 71010409 hypothetical protein UM02240.1 [Ustilago maydis 521] 17 XP_681529 67902546 hypothetical protein AN8260.2 [Aspergillus nidulans FGSC A4] 18 EAS37438 90307807 hypothetical protein CIMG_02792 [Coccidioides immitis RS] 19 XP_961026 85100788 lanosterol synthase related protein [MIPS] [Neurospora crassa OR74A] 20 XP_751627 70993560 oxidosqualene:lanosterol cyclase [Aspergillus fumigatus Af293] 21 EAQ86439 88178971 hypothetical protein CHGG_07692 [Chaetomium globosum CBS 148.51] 22 XP_386126 46123145 hypothetical protein FG05950.1 [Gibberella zeae PH-1] 23 BAE63415 83773288 unnamed protein product [Aspergillus oryzae] 24 EAS31167 90301536 hypothetical protein CIMG_06646 [Coccidioides immitis RS] 25 EAT81661 111060541 hypothetical protein SNOG_11162 [Phaeosphaeria nodorum SN15] 26 XP_570450 58266588 lanosterol synthase [Cryptococcus neoformans var.neoformans JEC21] 27 AAL56020 18028346 oxidosqualene:lanosterol cyclase [Cephalosporium caerulens] 28 BAC37102 26346907 unnamed protein product [Mus musculus] 29 Q8BLN5 62286881 Lanosterol synthase (Oxidosqualene--lanosterol cyclase) (2,3- epoxysqualene--lanosterol cyclase) (OSC) 30 XP_001 . . . 109065021 PREDICTED: lanosterol synthase isoform 2 [Macaca mulatta] 31 1W6KA 56966682 Chain A, Structure Of Human Osc In Complex With Lanosterol 32 BAC31739 26336108 unnamed protein product [Mus musculus] 33 XP_646246 66825783 hypothetical protein DDB_0191311 [Dictyostelium discoideum AX4] 34 AAF80384 8886139 cycloartenol synthase [Dictyostelium discoideum] 35 CAB42828 4808278 lanosterol synthase [Homo sapiens] 36 1W6JA 56966681 Chain A, Structure Of Human Osc In Complex With Ro 48-8071 37 BAA08208 639865 2,3-oxidosqualene:lanosterol cyclase [Rattus norvegicus] 38 NP_112311 13591981 lanosterol synthase [Rattus norvegicus] 39 NP_001 . . . 114053041 lanosterol synthase (2,3-oxidosqualene-lanosterol cyclase) [Bos taurus] 40 P48450 62296496 Lanosterol synthase (Oxidosqualene--lanosterol cyclase) (2,3- epoxysqualene--lanosterol cyclase) (OSC) 41 NP_001 . . . 57529773 lanosterol synthase (2,3-oxidosqualene-lanosterol cyclase) [Gallus gallus] 42 XP_751356 70993016 oxidosqualene cyclase [Aspergillus fumigatus Af293] 43 CAG08284 47222029 unnamed protein product [Tetraodon nigroviridis] 44 BAB83085 18147590 cycloartenol synthase [Betula platyphylla] 45 BAA76902 4589852 cycloartenol synthase [Glycyrrhiza glabra] 46 BAE61719 83771588 unnamed protein product [Aspergillus oryzae] 47 BAA33460 3688598 Cycloartenol Synthase [Panax ginseng] 48 XP_001 . . . 109065023 PREDICTED: lanosterol synthase isoform 1 [Macaca mulatta] 49 BAD34644 50896401 cycloartenol synthase [Cucurbita pepo] 50 BAA85266 6045133 cycloartenol synthase [Luffa aegyptiaca] 51 BAB83253 18147771 cycloartenol synthase [Costus speciosus] 52 ABB76767 82468805 cycloartenol synthase [Ricinus communis] 53 BAA23533 2627181 cycloartenol synthase [Pisum sativum] 54 BAD34645 50896403 cucurbitadienol synthase [Cucurbita pepo] 55 BAE53431 83016479 cycloartenol synthase [Lotus japonicus] 56 AAS83469 46242746 cycloartenol synthase [Bupleurum kaoi] 57 A49398 541855 cycloartenol synthase (EC 5.4.99.8) - Arabidopsis thaliana 58 AAN64509 24796994 At2g07050/T4E14.16 [Arabidopsis thaliana] 59 BAD34646 50896405 putative oxidosqualene cyclase [Cucurbita pepo] 60 AAC04931 452446 cycloartenol synthase; (S)-2,3-epoxysqualene mutase [Arabidopsis thaliana] 61 BAE95408 108743265 lanosterol synthase [Arabidopsis thaliana] 62 AAG44096 12004573 cycloartenol synthase [Abies magnifica] 63 AAS01524 41387168 cycloartenol synthase [Centella asiatica] 64 BAB83086 18147592 cycloartenol synthase [Betula platyphylla] 65 AAF03375 6090879 putative cycloartenol synthase [Oryza sativa] 66 BAF07762 113535379 Os02g0139700 [Oryza sativa (japonica cultivar-group)] 67 BAE95410 108743269 lanosterol synthase [Lotus japonicus] 68 AAK82995 15076959 lanosterol synthase [Trypanosoma cruzi] 69 XP_820967 71668047 lanosterol synthase [Trypanosoma cruzi strain CL Brener] 70 BAA85267 6045135 oxidosqualene cyclase [Luffa aegyptiaca] 71 NP_190099 42565553 catalytic/lyase [Arabidopsis thaliana] 72 AAT38889 47834383 cycloartenol synthase [Avena longiglumis] 73 BAA86933 6456469 oxidosqualene cyclase [Taraxacum officinale] 74 AAT38891 47834387 cycloartenol synthase [Avena strigosa] 75 CAC84559 15866702 cycloartenol synthase [Avena strigosa] 76 AAT38890 47834385 cycloartenol synthase [Avena prostrata] 77 AAT38888 47834381 cycloartenol synthase [Avena longiglumis] 78 AAT38887 47834379 cycloartenol synthase [Avena clauda] 79 AAT38892 47834389 cycloartenol synthase [Avena ventricosa] 80 XP_819758 71665580 lanosterol synthase [Trypanosoma cruzi strain CL Brener] 81 BAA86930 6456434 lupeol synthase [Olea europaea] 82 CAA61078 984145 lanosterol synthase [Homo sapiens] 83 BAA33462 3688602 Oxidosqualene Cyclase [Panax ginseng] 84 AAG26328 11023151 lanosterol synthase [Trypanosoma brucei brucei] 85 XP_825673 71752517 lanosterol synthase [Trypanosoma brucei TREU927] 86 BAA86932 6456467 lupeol synthase [Taraxacum officinale] 87 CAD23247 27475608 beta-amyrin synthase [Medicago truncatula] 88 BAA97558 8918271 beta-amyrin synthase [Pisum sativum] 89 BAA86931 6456465 cycloartenol synthase [Olea europaea] 90 BAD08587 41687978 lupeol synthase [Glycyrrhiza glabra] 91 BAA84603 5922599 oxidosqualene cyclase [Allium macrostemon] 92 AAO33578 28194504 beta-amyrin synthase [Medicago truncatula] 93 BAB83087 18147594 lupeol synthase [Betula platyphylla] 94 BAB83254 18147773 multifunctional triterpene synthase [Costus speciosus] 95 BAA89815 6730969 beta-amyrin synthase [Glycyrrhiza glabra] 96 BAE53429 83016474 beta-amyrin synthase [Lotus japonicus] 97 AAO33579 28194506 putative beta-amyrin synthase [Lotus japonicus] 98 BAA33461 3688600 beta-Amyrin Synthase [Panax ginseng] 99 CAB72151 6911851 oxidosqualene cyclase-like protein [Arabidopsis thaliana] 100 CAJ02110 68124127 lanosterol synthase, putative [Leishmania major] 101 BAA33722 3721856 beta-Amyrin Synthase [Panax ginseng] 102 AAX14716 60203059 beta-amyrin synthase [Aster sedifolius] 103 BAE53430 83016477 lupeol synthase [Lotus japonicus] 104 ABE91090 92892072 Prenyltransferase/squalene oxidase [Medicago truncatula] 105 XP_681518 67902524 hypothetical protein AN8249.2 [Aspergillus nidulans FGSC A4] 106 AAC17080 3152599 Strong similarity to lupeol synthase gb|U49919 and cycloartenol synthase gb|U02555 from A. thaliana (the third gene with similar homology). [Arabidopsis thaliana] 107 BAF28067 113644926 Os11g0286800 [Oryza sativa (japonica cultivar-group)] 108 NP_683508 22330736 beta-amyrin synthase [Arabidopsis thaliana] 109 AAS83468 46242744 beta-armyrin synthase [Bupleurum kaoi] 110 AAO33580 28194508 multifunctional beta-amyrin synthase [Lotus japonicus] 111 BAE43642 73991374 beta-amyrin synthase [Euphorbia tirucalli] 112 BAB83088 18147596 beta-amyrin synthase [Betula platyphylla] 113 ABB76766 82468803 lupeol synthase [Ricinus communis] 114 ABG22399 108864084 Cycloartenol synthase, putative, expressed [Oryza sativa (japonica cultivar- group)] 115 BAF28063 113644922 Os11g0285000 [Oryza sativa (japonica cultivar-group)] 116 ABG22398 108864083 Cycloartenol synthase, putative, expressed [Oryza sativa (japonica cultivar- group)] 117 NP_849903 30699380 LUP1 (LUPEOL SYNTHASE 1); lupeol synthase [Arabidopsis thaliana] 118 AAB94341 2738027 2,3-oxidosqualene-triterpenoid cyclase [Arabidopsis thaliana] 119 AAN77001 25991999 2,3-oxidosqualene-triterpene cyclase [Arabidopsis thaliana] 120 AAT38893 47834391 beta-amyrin synthase [Avena clauda] 121 AAD05032 1762150 lupeol synthase [Arabidopsis thaliana] 122 NP_178017 30699377 ATLUP2; lupeol synthase [Arabidopsis thaliana] 123 ABE88599 92887080 squalene/oxidosqualene cyclases [Medicago truncatula] 124 ABG22449 108864253 Cycloartenol synthase, putative, expressed [Oryza sativa (japonica cultivar- group)] 125 BAB68529 15787841 isomultiflorenol synthase [Luffa aegyptiaca] 126 AAT38898 47834401 beta-amyrin synthase [Avena ventricosa] 127 AAT38894 47834393 beta-amyrin synthase [Avena longiglumis] 128 AAT38897 47834399 beta-amyrin synthase [Avena strigosa] 129 NP_178016 22330734 catalytic/lyase [Arabidopsis thaliana] 130 AAT38895 47834395 beta-amyrin synthase [Avena longiglumis] 131 AAT38896 47834397 beta-amyrin synthase [Avena prostrata] 132 BAA97559 8918273 mixed-amyrin synthase [Pisum sativum] 133 AAC98864 4090722 pentacyclic triterpene synthase [Arabidopsis thaliana] 134 BAF28466 113645325 Os11g0562100 [Oryza sativa (japonica cultivar-group)] 135 AAG41762 11934652 pentacyclic triterpene synthase [synthetic construct] 136 XP_480759 50942463 putative Cycloartenol Synthase [Oryza sativa (japonica cultivar-group)] 137 BAE43643 73991380 putative oxidosqulene cyclase [Euphorbia tirucalli] 138 AAC17070 3152589 Strong similarity to lupeol synthase gb|U49919 from A. thaliana (second gene in a series of three with similar homologies). [Arabidopsis thaliana] 139 BAD15332 46359655 beta-amyrin synthase [Panax ginseng] 140 AAM23264 23428800 beta-amyrin synthase [Glycine max] 141 ABC33922 83638481 beta-amyrin synthase [Gypsophila paniculata] 142 AAS01523 41387158 putative beta-amyrin synthase [Centella asiatica] 143 BAE99758 110743847 cycloartenol synthase [Arabidopsis thaliana] 144 BAB11065 10177752 cycloartenol synthase [Arabidopsis thaliana] 145 NP_177971 15218390 catalytic [Arabidopsis thaliana] 146 CAD39196 32526539 cycloartenol synthase [Stigmatella aurantiaca] 147 ABG22450 108864254 Cycloartenol synthase, putative, expressed [Oryza sativa (japonica cultivar- group)] 148 CAA93571 1204221 erg7 [Schizosaccharomyces pombe] 149 NP_567462 18414430 ATPEN1; catalytic/lyase [Arabidopsis thaliana] 150 NP_199612 42568386 catalytic [Arabidopsis thaliana] 151 NP_193272 15233798 catalytic/lyase [Arabidopsis thaliana] 152 ABE88610 92887091 squalene/oxidosqualene cyclases [Medicago truncatula] 153 AAW30034 56790234 At5g42600 [Arabidopsis thaliana] 154 BAF01935 110740066 putative lupeol synthase [Arabidopsis thaliana] 155 NP_198464 15239312 ATPEN3; catalytic [Arabidopsis thaliana] 156 BAB83089 18147598 putative oxidosqualene cyclase [Betula platyphylla] 157 YP_115266 53803023 squalene cyclase family protein [Methylococcus capsulatus str. Bath] 158 CAB78579 7268284 lupeol synthase like protein [Arabidopsis thaliana] 159 CAB78576 7268281 lupeol synthase like protein [Arabidopsis thaliana]

TABLE 56 Examples of ERG6 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 CAA89944 854482 Erg6p [Saccharomyces cerevisiae] 2 P25087 462024 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 3 CAA52308 396515 S-adenosyl-methionine:delta-24-sterol-C-methyltransferase [Saccharomyces cerevisiae] 4 AAB31378 619251 putative S-adenosylmethionine-dependent methyltransferase [Saccharomyces cerevisiae] 5 Q6FRZ7 62900214 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 6 AAX73200 62178560 C24 sterol methyltransferase [Candida glabrata] 7 P_446997 50289135 unnamed protein product [Candida glabrata] 8 CAG59930 49526306 unnamed protein product [Candida glabrata CBS138] 9 AAX73199 62178558 C24 sterol methyltransferase [Candida glabrata] 10 Q6CYB3 62900210 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 11 P_451076 50302283 unnamed protein product [Kluyveromyces lactis] 12 CAH02664 49640207 unnamed protein product [Kluyveromyces lactis NRRL Y-1140] 13 Q759S7 62900245 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 14 P_984292 45188069 ADR196Wp [Eremothecium gossypii] 15 AAS52116 44982886 ADR196Wp [Ashbya gossypii ATCC 10895] 16 CAA37826 4122 unnamed protein product [Saccharomyces cerevisiae] 17 Q875K1 62900279 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 18 AAO21936 27803704 S-adenosylmethionine:D24-methyltransferase [Clavispora lusitaniae] 19 Q6BRB7 62900203 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 20 P_459253 50421405 hypothetical protein DEHA0D19151g [Debaryomyces hansenii CBS767] 21 CAG87427 49654920 unnamed protein product [Debaryomyces hansenii CBS767] 22 P_721588 68468538 SAM:delta 24-methyltransferase [Candida albicans SC5314] 23 P_721708 68468297 SAM:delta 24-methyltransferase [Candida albicans SC5314] 24 EAL02920 46443640 hypothetical protein CaO19.1631 [Candida albicans SC5314] 25 EAL02792 46443511 hypothetical protein CaO19.9199 [Candida albicans SC5314] 26 O74198 6015114 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 27 AAC26626 3323500 sterol transmethylase [Candida albicans] 28 Q6C2D9 62900205 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 29 P_505173 50555530 hypothetical protein [Yarrowia lipolytica] 30 CAG77980 49651043 unnamed protein product [Yarrowia lipolytica CLIB122] 31 P_595787 19112579 hypothetical protein SPBC16E9.05 [Schizosaccharomyces pombe 972h-] 32 O14321 6166151 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 33 CAB16897 2467267 SPBC16E9.05 [Schizosaccharomyces pombe] 34 EAT84719 111063599 hypothetical protein SNOG_08443 [Phaeosphaeria nodorum SN15] 35 EAS29698 90300067 hypothetical protein CIMG_08444 [Coccidioides immitis RS] 36 EAQ70709 86196071 hypothetical protein MGG_ch7g116 [Magnaporthe grisea 70-15] 37 P_366350 39969919 hypothetical protein MG10568.4 [Magnaporthe grisea 70-15] 38 P_746550 70982043 sterol 24-c-methyltransferase [Aspergillus fumigatus Af293] 39 EAL84512 66844173 sterol 24-c-methyltransferase, putative [Aspergillus fumigatus Af293] 40 BAE64809 83774686 unnamed protein product [Aspergillus oryzae] 41 P_965392 85118140 probable DELTA(24)-STEROL C-METHYLTRANSFERASE [MIPS] [Neurospora crassa OR74A] 42 Q9P3R1 62900336 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 43 EAA36156 28927200 probable DELTA(24)-STEROL C-METHYLTRANSFERASE [MIPS] [Neurospora crassa] 44 CAB97289 9367272 probable DELTA(24)-STEROL C-METHYLTRANSFERASE (ERG6) [Neurospora crassa] 45 P_664750 67541965 hypothetical protein AN7146.2 [Aspergillus nidulans FGSC A4] 46 EAA61398 40742208 hypothetical protein AN7146.2 [Aspergillus nidulans FGSC A4] 47 P_382959 46111803 hypothetical protein FG02783.1 [Gibberella zeae PH-1] 48 EAA70778 42547935 hypothetical protein FG02783.1 [Gibberella zeae PH-1] 49 Q96WX4 62900323 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 50 AAK54439 14192727 S-adenosyl methionine:sterol methyl transferase [Pneumocystis carinii] 51 EAQ89522 88182054 conserved hypothetical protein [Chaetomium globosum CBS 148.51] 52 BAA13793 41688402 unnamed protein product [Schizosaccharomyces pombe] 53 P_385916 46122725 hypothetical protein FG05740.1 [Gibberella zeae PH-1] 54 EAA75815 42552972 hypothetical protein FG05740.1 [Gibberella zeae PH-1] 55 P_361872 39944670 hypothetical protein MG04346.4 [Magnaporthe grisea 70-15] 56 Q5EN22 62900141 Sterol 24-C-methyltransferase (Delta(24)-sterol C- methyltransferase) 57 AAX07631 59802827 sterol 24-C-methyltransferase-like protein [Magnaporthe grisea] 58 BAE61033 83770900 unnamed protein product [Aspergillus oryzae] 59 P_759329 71018197 hypothetical protein UM03182.1 [Ustilago maydis 521] 60 EAK84412 46099179 hypothetical protein UM03182.1 [Ustilago maydis 521] 61 AAK00294 12642580 sterol methyl transferase [Pneumocystis carinii f. sp. carinii] 62 P_360548 39942022 hypothetical protein MG10860.4 [Magnaporthe grisea 70-15] 63 P_568887 58262954 sterol 24-C-methyltransferase [Cryptococcus neoformans var. neoformans JEC21] 64 AAW41580 57223537 sterol 24-C-methyltransferase, putative [Cryptococcus neoformans var. neoformans JEC21] 65 EAL22628 50259962 hypothetical protein CNBB2600 [Cryptococcus neoformans var. neoformans B-3501A] 66 AAC34951 3560474 S-adenosyl-methionine-sterol-C-methyltransferase [Nicotiana tabacum] 67 AAC04265 2909846 (S)-adenosyl-L-methionine:delta 24-sterol methyltransferase [Zea mays] 68 AAB49338 1872473 delta-24-sterol methyltransferase [Triticum aestivum] 69 Q9LM02 62901053 Cycloartenol-C-24-methyltransferase (24-sterol C-methyltransferase 1) (Sterol C-methyltransferase 1) (Protein STEROL METHYLTRANSFERASE 1) (Protein CEPHALOPOD) 70 AAN15377 23197700 24-sterol C-methyltransferase [Arabidopsis thaliana] 71 AAM53274 21539443 24-sterol C-methyltransferase [Arabidopsis thaliana] 72 P_196875 15240691 SMT1 (STEROL METHYLTRANSFERASE 1) [Arabidopsis thaliana] 73 AAG28462 11066105 sterol methyltransferase SMT1 [Arabidopsis thaliana] 74 BAB08698 9758037 24-sterol C-methyltransferase [Arabidopsis thaliana] 75 AAF78847 8745241 SAM:cycloartenol-C24-methyltransferase [Arabidopsis thaliana] 76 AAM53553 21434843 cephalopod [Arabidopsis thaliana] 77 AAB70886 1899060 endosperm C-24 sterol methyltransferase [Zea mays] 78 AAB37769 1706965 delta-24-sterol methyltransferase [Triticum aestivum] 79 BAF21068 113610690 Os07g0206700 [Oryza sativa (japonica cultivar-group)] 80 Q6ZIX2 68566037 Cycloartenol-C-24-methyltransferase 1 (24-sterol C- methyltransferase 1) (Sterol C-methyltransferase 1) 81 P_477078 50935101 cycloartenol-C24-methyltransferase [Oryza sativa (japonica cultivar-group)] 82 BAC83238 34393309 cycloartenol-C24-methyltransferase [Oryza sativa (japonica cultivar-group)] 83 AAB04057 1399380 S-adenosyl-L-methionine:delta24-sterol-C-methyltransferase 84 AAB62812 2246458 S-adenosyl-methionine-sterol-C-methyltransferase [Ricinus communis] 85 AAC35787 3603295 S-adenosyl-methionine cycloartenol-C24-methyltransferase [Nicotiana tabacum] 86 AAC34988 3560531 cycloartenol-C24-methyltransferase [Oryza sativa subsp. japonica] 87 AAZ83345 73761691 24-sterol C-methyltransferase [Gossypium hirsutum] 88 EAS30660 90301029 hypothetical protein CIMG_06139 [Coccidioides immitis RS] 89 P_470035 50919277 putative endosperm C-24 sterol methyltransferase [Oryza sativa (japonica cultivar-group)] 90 AAP21419 30103006 putative endosperm C-24 sterol methyltransferase [Oryza sativa (japonica cultivar-group)] 91 AAR92099 40806291 S-adenosyl-L-methionine-C-24-delta-sterol-methyltransferase B [Leishmania donovani] 92 AAR92098 40806289 S-adenosyl-L-methionine-C-24-delta-sterol-methyltransferase A [Leishmania donovani] 93 CAJ09197 68129891 sterol 24-c-methyltransferase, putative [Leishmania major] 94 CAJ09196 68129890 sterol 24-c-methyltransferase, putative [Leishmania major] 95 P_802864 71399753 sterol 24-c-methyltransferase [Trypanosoma cruzi strain CL Brener] 96 EAN81418 70865128 sterol 24-c-methyltransferase, putative [Trypanosoma cruzi] 97 P_802716 71399140 sterol 24-c-methyltransferase [Trypanosoma cruzi strain CL Brener] 98 EAN81270 70864663 sterol 24-c-methyltransferase, putative [Trypanosoma cruzi] 99 P_822930 71747750 sterol 24-c-methyltransferase [Trypanosoma brucei TREU927] 100 AAZ40214 71738257 sterol methyltransferase [Trypanosoma brucei brucei] 101 CAJ16958 70908212 s-adenosyl-L-methionine-c-24-delta-sterol-methyltransferase a, putative [Trypanosoma brucei] 102 EAN78102 70832598 sterol 24-c-methyltransferase, putative [Trypanosoma brucei] 103 P_636481 66805519 hypothetical protein DDBDRAFT_0188166 [Dictyostelium discoideum AX4] 104 EAL62977 60464861 hypothetical protein DDBDRAFT_0188166 [Dictyostelium discoideum AX4] 105 ABF99454 108711659 Cyclopropane-fatty-acyl-phospholipid synthase family protein, expressed [Oryza sativa (japonica cultivar-group)] 106 P_802811 71399535 sterol 24-c-methyltransferase [Trypanosoma cruzi strain CL Brener] 107 EAN81365 70864963 sterol 24-c-methyltransferase, putative [Trypanosoma cruzi] 108 ABB90541 82780739 c24-sterol methyltransferase [Paracoccidioides brasiliensis] 109 AAB62807 2246452 S-adenosyl-methionine-sterol-C-methyltransferase homolog [Nicotiana tabacum] 110 Q39227 62900893 24-methylenesterol C-methyltransferase 2 (24-sterol C- methyltransferase 2) (Sterol-C-methyltransferase 2) 111 AAM45009 21281107 putative sterol-C-methyltransferase [Arabidopsis thaliana] 112 P_173458 15217917 SMT2 (STEROL METHYLTRANSFERASE 2) [Arabidopsis thaliana] 113 AAK76716 15028255 putative sterol-C-methyltransferase [Arabidopsis thaliana] 114 AAG48780 12083242 putative sterol-C-methyltransferase [Arabidopsis thaliana] 115 AAF88156 9558593 Identical to 24-sterol C-methyltransferase from Arabidopsis thaliana gi|2129517 and is a member of the ubiE/COQ5 methyltransferase family PF|01209. ESTs gb|T42228, gb|T46520, gb|T41746, gb|N38458, gb|AI993515, gb|AA389843, gb|AI099890, gb|AI099653 come f 116 2207220A 1587694 sterol C-methyltransferase 117 CAA61966 1061040 sterol-C-methyltransferase [Arabidopsis thaliana] 118 AAN72104 25083682 24-sterol C-methyltransferase [Arabidopsis thaliana] 119 AAM91592 22136020 24-sterol C-methyltransferase [Arabidopsis thaliana] 120 Q94JS4 67472706 24-methylenesterol C-methyltransferase 3 (24-sterol C- methyltransferase 3) (Sterol-C-methyltransferase 3) 121 AAM47339 21360447 At1g76090/T23E18_40 [Arabidopsis thaliana] 122 P_177736 15222955 SMT3; S-adenosylmethionine-dependent methyltransferase [Arabidopsis thaliana] 123 AAK52981 14030613 At1g76090/T23E18_40 [Arabidopsis thaliana] 124 AAN31890 23397216 putative sterol-C-methyltransferase [Arabidopsis thaliana] 125 AAB62809 2246456 S-adenosyl-methionine-sterol-C-methyltransferase [Arabidopsis thaliana] 126 AAB62808 2246454 S-adenosyl-methionine-sterol-C-methyltransferase [Nicotiana tabacum] 127 AAM63753 21555010 sterol-C-methyltransferase [Arabidopsis thaliana] 128 BAF10805 113547362 Os03g0136200 [Oryza sativa (japonica cultivar-group)] 129 ABF93854 108706059 24-methylenesterol C-methyltransferase 2, putative, expressed [Oryza sativa (japonica cultivar-group)] 130 O82427 68566013 24-methylenesterol C-methyltransferase 2 (24-sterol C- methyltransferase 2) (Sterol-C-methyltransferase 2)

TABLE 57 Examples of ERG5 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 P54781 1706693 Cytochrome P450 61 (C-22 sterol desaturase) 2 AAB06217 1235991 cytochrome P450 3 CAA89116 798924 unknown [Saccharomyces cerevisiae] 4 AAU09769 51830482 YMR015C [Saccharomyces cerevisiae] 5 AAX73198 62178556 C22 sterol desaturase [Candida glabrata] 6 AAX73197 62178554 C22 sterol desaturase [Candida glabrata] 7 P_449681 50294540 hypothetical protein CAGL0M07656g [Candida glabrata CBS138] 8 CAG62657 49528995 unnamed protein product [Candida glabrata CBS138] 9 P_453562 50307167 unnamed protein product [Kluyveromyces lactis] 10 CAH00658 49642696 unnamed protein product [Kluyveromyces lactis NRRL Y-1140] 11 P_982999 45185282 ABR053Cp [Eremothecium gossypii] 12 AAS50823 44980940 ABR053Cp [Ashbya gossypii ATCC 10895] 13 P_458727 50420383 hypothetical protein DEHA0D06842g [Debaryomyces hansenii CBS767] 14 CAG86871 49654394 unnamed protein product [Debaryomyces hansenii CBS767] 15 P_888893 77022898 hypothetical protein CaJ7.0323 [Candida albicans SC5314] 16 BAE44790 76573706 hypothetical protein [Candida albicans] 17 P_716933 68478128 cytochrome P450 [Candida albicans SC5314] 18 P_717000 68477993 hypothetical protein CaO19.5178 [Candida albicans SC5314] 19 EAK98020 46438693 hypothetical protein CaO19.5178 [Candida albicans SC5314] 20 EAK97950 46438622 hypothetical protein CaO19.12645 [Candida albicans SC5314] 21 CAA21953 3850153 cytochrome P450 [Candida albicans] 22 P_500188 50545301 hypothetical protein [Yarrowia lipolytica] 23 CAG84120 49646053 unnamed protein product [Yarrowia lipolytica CLIB122] 24 BAC01142 21624361 sterol C-22 desaturase [Symbiotaphrina buchneri] 25 BAC01140 21624354 sterol C-22 desaturase [Symbiotaphrina buchneri] 26 BAC01141 21624359 sterol C-22 desaturase [Symbiotaphrina kochii] 27 BAC01139 21624352 sterol C22 desaturase [Symbiotaphrina kochii] 28 EAQ88206 88180738 conserved hypothetical protein [Chaetomium globosum CBS148.51] 29 P_961915 85105220 hypothetical protein [Neurospora crassa OR74A] 30 EAA32679 28923500 hypothetical protein [Neurospora crassa] 31 EAS33985 90304354 hypothetical protein CIMG_05009 [Coccidioides immitis RS] 32 P_750145 70990592 cytochrome P450 sterol C-22 desaturase [Aspergillus fumigatus Af293] 33 EAL88107 66847777 cytochrome P450 sterol C-22 desaturase, putative [Aspergillus fumigatus Af293] 34 EAT87119 111065999 hypothetical protein SNOG_06055 [Phaeosphaeria nodorum SN15] 35 BAE58068 83767929 unnamed protein product [Aspergillus oryzae] 36 P_382135 46110154 hypothetical protein FG01959.1 [Gibberella zeae PH-1] 37 EAA68855 42546012 hypothetical protein FG01959.1 [Gibberella zeae PH-1] 38 P_661646 67527317 hypothetical protein AN4042.2 [Aspergillus nidulans FGSC A4] 39 EAA59513 40740323 hypothetical protein AN4042.2 [Aspergillus nidulans FGSC A4] 40 P_383862 46115688 hypothetical protein FG03686.1 [Gibberella zeae PH-1] 41 EAA71499 42548656 hypothetical protein FG03686.1 [Gibberella zeae PH-1] 42 P_369417 39976053 hypothetical protein MG06047.4 [Magnaporthe grisea 70-15] 43 BAE57461 83767322 unnamed protein product [Aspergillus oryzae] 44 P_593788 19114700 hypothetical protein SPAC19A8.04 [Schizosaccharomyces pombe 972h-] 45 CAB11640 7007345 SPAC19A8.04 [Schizosaccharomyces pombe] 46 P_756497 71003662 hypothetical protein UM00350.1 [Ustilago maydis 521] 47 EAK81168 46095935 hypothetical protein UM00350.1 [Ustilago maydis 521] 48 P_571445 58268578 C-22 sterol desaturase [Cryptococcus neoformans var. neoformans JEC21] 49 AAW44138 57227680 C-22 sterol desaturase, putative [Cryptococcus neoformans var. neoformans JEC21] 50 EAL20298 50257593 hypothetical protein CNBF1100 [Cryptococcus neoformans var. neoformans B-3501A] 51 EAQ86140 88178672 hypothetical protein CHGG_07393 [Chaetomium globosum CBS 148.51] 52 AAO48601 28864257 ERG5 [Clavispora lusitaniae] 53 P_647018 66827327 cytochrome P450 family protein [Dictyostelium discoideum AX4] 54 EAL73097 60475161 cytochrome P450 family protein [Dictyostelium discoideum AX4] 55 BAF04287 113531904 Os01g0211200 [Oryza sativa (japonica cultivar-group)] 56 P_913287 34903880 unnamed protein product [Oryza sativa (japonica cultivar-group)] 57 BAA96196 8096624 putative sterol C-22 desaturase [Oryza sativa (japonica cultivar- group)] 58 BAA96154 8096581 putative sterol C-22 desaturase [Oryza sativa (japonica cultivar- group)] 59 ABC59097 84514177 cytochrome P450 monooxygenase CYP710A15 [Medicago truncatula] 60 P_180452 15226976 CYP710A4; heme binding/iron ion binding/monooxygenase/ oxygen binding [Arabidopsis thaliana] 61 AAC79590 3927833 putative cytochrome P450 [Arabidopsis thaliana] 62 P_913285 34903876 unnamed protein product [Oryza sativa (japonica cultivar-group)] 63 BAA96194 8096622 sterol C-22 desaturase-like [Oryza sativa (japonica cultivar-group)] 64 BAA96152 8096579 sterol C-22 desaturase-like [Oryza sativa (japonica cultivar-group)] 65 P_913291 34903888 unnamed protein product [Oryza sativa (japonica cultivar-group)] 66 BAA96200 8096628 sterol C-22 desaturase-like [Oryza sativa (japonica cultivar-group)] 67 BAA96158 8096585 sterol C-22 desaturase-like [Oryza sativa (japonica cultivar-group)] 68 BAF04286 113531903 Os01g0210900 [Oryza sativa (japonica cultivar-group)] 69 P_913284 34903874 unnamed protein product [Oryza sativa (japonica cultivar-group)] 70 BAA96193 8096621 sterol C-22 desaturase-like [Oryza sativa (japonica cultivar-group)] 71 BAA96151 8096578 sterol C-22 desaturase-like [Oryza sativa (japonica cultivar-group)] 72 BAE71351 84468537 CYP710 [Arabidopsis thaliana] 73 AAQ65177 34365731 At2g34500 [Arabidopsis thaliana] 74 AAN72080 25083451 putative cytochrome P450 [Arabidopsis thaliana] 75 AAM14944 20197156 putative cytochrome P450 [Arabidopsis thaliana] 76 P_180997 15226758 CYP710A1; heme binding/iron ion binding/monooxygenase/ oxygen binding [Arabidopsis thaliana] 77 AAC26690 3128210 putative cytochrome P450 [Arabidopsis thaliana] 78 P_180451 15226974 CYP710A3; heme binding/iron ion binding/monooxygenase/ oxygen binding [Arabidopsis thaliana] 79 AAC79589 3927832 putative cytochrome P450 [Arabidopsis thaliana] 80 BAE80316 89111285 CYP710 protein [Arabidopsis thaliana] 81 AAM26664 20856398 At2g34490/T31E10.17 [Arabidopsis thaliana] 82 AAM14945 20197157 putative cytochrome P450 [Arabidopsis thaliana] 83 AAL75888 18650637 At2g34490/T31E10.17 [Arabidopsis thaliana] 84 P_180996 15226756 CYP710A2; heme binding/iron ion binding/monooxygenase/ oxygen binding [Arabidopsis thaliana] 85 AAC26691 3128211 putative cytochrome P450 [Arabidopsis thaliana] 86 AAM62799 21553706 putative cytochrome P450 [Arabidopsis thaliana] 87 BAE93156 91204699 sterol 22-desaturase [Lycopersicon esculentum]

TABLE 58 Examples of ERG3 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAT93015 51013443 YLR056W [Saccharomyces cerevisiae] 2 NP_013157 6323085 C-5 sterol desaturase, catalyzes the introduction of a C-5(6) 3 BAA20292 2143190 C-5 sterol desaturase [Saccharomyces cerevisiae] 4 CAA64303 1181277 C-5 sterol desaturase [Saccharomyces cerevisiae] 5 P32353 416963 C-5 sterol desaturase (Sterol-C5-desaturase) (Ergosterol delta 6 AAB39844 233331 C-5 sterol desaturase [Saccharomyces cerevisiae] 7 AAA34595 171477 C-5 sterol desaturase 8 AAA34594 171475 C-5 sterol desaturase 9 XP_446050 50287241 unnamed protein product [Candida glabrata] 10 CAG58974 49525357 unnamed protein product [Candida glabrata CBS138] 11 P50860 1706691 C-5 sterol desaturase (Sterol-C5-desaturase) (Ergosterol delta 12 AAB02330 755695 ERG3 13 Q754B9 51701406 C-5 sterol desaturase (Sterol-C5-desaturase) (Ergosterol delta 14 NP_985698 45198669 AFR151Cp [Eremothecium gossypii] 15 AAS53522 44984679 AFR151Cp [Ashbya gossypii ATCC 10895A] 16 XP_455482 50310919 unnamed protein product [Kluyveromyces lactis] 17 CAG98190 49644618 unnamed protein product [Kluyveromyces lactis NRRL Y-1140] 18 Q8NJ57 51701432 C-5 sterol desaturase (Sterol-C5-desaturase) (Ergosterol delta 19 CAD13131 22022945 ergosterol delta 5,6 desaturase [Candida dubliniensis] 20 XP_713577 68485022 hypothetical protein CaO19.767 [Candida albicans SC5314] 21 XP_713612 68484947 hypothetical protein CaO19.8387 [Candida albicans SC5314] 22 EAK94507 46435118 hypothetical protein CaO19.8387 [Candida albicans SC5314] 23 EAK94472 46435082 hypothetical protein CaO19.767 [Candida albicans SC5314] 24 O93875 51701379 C-5 sterol desaturase (Sterol-C5-desaturase) (Ergosterol delta 25 AAC99343 4091929 C5,6 desaturase [Candida albicans] 26 XP_460747 50424321 hypothetical protein DEHA0F09757g [Debaryomyces hansenii CBS76 27 CAG89088 49656416 unnamed protein product [Debaryomyces hansenii CBS767] 28 XP_503090 50551233 hypothetical protein [Yarrowia lipolytica] 29 CAG81282 49648958 unnamed protein product [Yarrowia lipolytica CLIB122] 30 XP_664110 67540672 hypothetical protein AN6506.2 [Aspergillus nidulans FGSC A4] 31 EAA57846 40738656 hypothetical protein AN6506.2 [Aspergillus nidulans FGSC A4] 32 BAE61784 83771654 unnamed protein product [Aspergillus oryzae] 33 XP_747563 70984086 sterol delta 5,6-desaturase ERG3 [Aspergillus fumigatus Af293] 34 EAL85525 66845190 sterol delta 5,6-desaturase ERG3 [Aspergillus fumigatus Af293] 35 AAU82098 52548218 C-5 sterol desaturase B-like [Aspergillus fumigatus] 36 BAE56076 83765933 unnamed protein product [Aspergillus oryzae] 37 XP_962923 85109451 hypothetical protein [Neurospora crassa OR74A] 38 Q7SBB6 51701418 Probable C-5 sterol desaturase (Sterol-C5-desaturase) (Ergoste 39 EAA33687 28924567 hypothetical protein [Neurospora crassa] 40 EAS37256 90307625 C-5 sterol desaturase [Coccidioides immitis RS] 41 Q8J207 51701428 C-5 sterol desaturase (Sterol-C5-desaturase) (Ergosterol delta 42 AAN27998 23476431 sterol delta 5,6-desaturase ERG3 [Leptosphaeria maculans] 43 XP_363987 39953467 hypothetical protein MG08832.4 [Magnaporthe grisea 70-15] 44 EAT83541 111062421 hypothetical protein SNOG_09349 [Phaeosphaeria nodorum SN15] 45 O94457 51701380 Probable C-5 sterol desaturase 1 (Sterol-C5-desaturase 1) (Erg 46 NP_593135 19114047 hypothetical protein SPAC1687.16c [Schizosaccharomyces pombe 9 47 CAA22610 4106670 SPAC1687.16c [Schizosaccharomyces pombe] 48 XP_382678 46111241 hypothetical protein FG02502.1 [Gibberella zeae PH-1] 49 EAA67236 42544393 hypothetical protein FG02502.1 [Gibberella zeae PH-1] 50 BAE65607 83775487 unnamed protein product [Aspergillus oryzae] 51 O13666 6015113 Probable C-5 sterol desaturase 2 (Sterol-C5-desaturase 2) (Erg 52 CAA16898 2853109 SPBC27B12.03c [Schizosaccharomyces pombe] 53 BAA21457 2257565 C-5 STEROL DESATURASE [Schizosaccharomyces pombe]

TABLE 59 Examples of ERG2 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAS56448 45270134 YMR202W [Saccharomyces cerevisiae] 2 CAA88643 736299 Erg2p [Saccharomyces cerevisiae] 3 P32352 416962 C-8 sterol isomerase (Delta-8--delta-7 sterol isomerase) 4 AAA34593 171473 C-8 sterol isomerase 5 P_985369 45198340 AFL181Cp [Eremothecium gossypii] 6 AAS53193 44984227 AFL181Cp [Ashbya gossypii ATCC 10895] 7 P_449233 50293643 unnamed protein product [Candida glabrata] 8 CAG62207 49528546 unnamed protein product [Candida glabrata CBS138] 9 P_455367 50310691 unnamed protein product [Kluyveromyces lactis] 10 CAG98075 49644503 unnamed protein product [Kluyveromyces lactis NRRL Y-1140] 11 P_458891 50420709 hypothetical protein DEHA0D10714g [Debaryomyces hansenii CBS767] 12 CAG87043 49654558 unnamed protein product [Debaryomyces hansenii CBS767] 13 P_718886 68473979 sterol C8-C7 isomerase [Candida albicans SC5314] 14 P_718988 68473770 sterol C8-C7 isomerase [Candida albicans SC5314] 15 EAL00087 46440785 hypothetical protein CaO19.6026 [Candida albicans SC5314] 16 EAK99982 46440679 hypothetical protein CaO19.13447 [Candida albicans SC5314] 17 P_504668 50554519 hypothetical protein [Yarrowia lipolytica] 18 CAG80272 49650537 unnamed protein product [Yarrowia lipolytica CLIB122] 19 P_501006 50547073 hypothetical protein [Yarrowia lipolytica] 20 CAG83259 49646872 unnamed protein product [Yarrowia lipolytica CLIB122] 21 Q92254 67476925 C-8 sterol isomerase (Delta-8--delta-7 sterol isomerase) 22 CAC28749 12718205 C-8 sterol isomerase erg-1 [Neurospora crassa] 23 AAB09470 1575320 C-8 sterol isomerase 24 P_658055 67516339 hypothetical protein AN0451.2 [Aspergillus nidulans FGSC A4] 25 EAA66550 40747394 hypothetical protein AN0451.2 [Aspergillus nidulans FGSC A4] 26 P_593324 19114236 hypothetical protein SPAC20G8.07c [Schizosaccharomyces pombe 972h-] 27 P87113 3219796 Probable C-8 sterol isomerase (Delta-8--delta-7 sterol isomerase) 28 CAB08601 2094862 SPAC20G8.07c [Schizosaccharomyces pombe] 29 P_363040 39947737 hypothetical protein MG08624.4 [Magnaporthe grisea 70-15] 30 P33281 462023 C-8 sterol isomerase (Delta-8--delta-7 sterol isomerase) 31 CAA80454 311322 C-8 sterol isomerase [Magnaporthe grisea] 32 EAQ85888 88178420 hypothetical protein CHGG_07141 [Chaetomium globosum CBS148.51] 33 P_961312 85102300 C-8 STEROL ISOMERASE (DELTA-8--DELTA-7 STEROL ISOMERASE) [Neurospora crassa OR74A] 34 EAA32076 28922855 C-8 STEROL ISOMERASE (DELTA-8--DELTA-7 STEROL ISOMERASE) [Neurospora crassa] 35 P_844057 72387265 C-8 sterol isomerase, putative [Trypanosoma brucei] 36 P_826540 71905267 C-8 sterol isomerase, putative [Trypanosoma brucei] 37 AAZ10498 70800589 C-8 sterol isomerase, putative [Trypanosoma brucei] 38 AAX80634 62360216 C-8 sterol isomerase, putative [Trypanosoma brucei] 39 P_758081 71006989 C-8 sterol isomerase [Ustilago maydis 521] 40 EAK82388 46097155 ERG2_USTMA C-8 sterol isomerase (Delta-8--delta-7 sterol isomerase) [Ustilago maydis 521] 41 P32360 416961 C-8 sterol isomerase (Delta-8--delta-7 sterol isomerase) 42 CAA78959 2967 C-8 sterol isomerase [Ustilago maydis] 43 P_567070 58259315 C-8 sterol isomerase [Cryptococcus neoformans var. neoformans JEC21] 44 AAW41251 57223207 C-8 sterol isomerase, putative [Cryptococcus neoformans var. neoformans JEC21] 45 EAL23045 50260386 hypothetical protein CNBA8120 [Cryptococcus neoformans var. neoformans B-3501A] 46 P_821186 71658920 C-8 sterol isomerase [Trypanosoma cruzi strain CL Brener] 47 EAN99335 70886558 C-8 sterol isomerase, putative [Trypanosoma cruzi] 48 AAQ18796 33439494 C-8 sterol isomerase [Trypanosoma cruzi] 49 P_847879 73536920 C-8 sterol isomerase-like protein [Leishmania major strain Friedlin] 50 AAZ09671 70799755 C-8 sterol isomerase-like protein [Leishmania major strain Friedlin]

TABLE 60 Examples of ERG3-like polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 BAA33730 3721884 sterol-C5-desaturase [Mus musculus] 2 BAE32506 74178504 unnamed protein product [Mus musculus] 3 AAH24132 18848246 Sterol-C5-desaturase (fungal ERG3, delta-5-desaturase) homolog (S. cerevisae) [Mus musculus] 4 NP_766357 27777693 sterol-C5-desaturase (fungal ERG3, delta-5-desaturase) homolog [Mus musculus] 5 BAC36944 26346591 unnamed protein product [Mus musculus] 6 BAC31666 26335931 unnamed protein product [Mus musculus] 7 AAH81704 51859106 Sterol-C5-desaturase (fungal ERG3, delta-5-desaturase) homolog (S. cerevisae) [Rattus norvegicus 8 NP_446094 25742651 sterol-C5-desaturase (fungal ERG3, delta-5-desaturase) homolog [Rattus norvegicus] 9 BAB19798 11990472 sterol C5-desaturase [Rattus norvegicus] 10 XP_853004 73955000 PREDICTED: similar to sterol-C5-desaturase-like [Canis familiaris] 11 XP_508825 55637121 PREDICTED: similar to delta7-sterol-C5-desaturase [Pan troglodytes] 12 NP_001 . . . 68160945 sterol-C5-desaturase-like [Homo sapiens] 13 NP_008849 68160941 sterol-C5-desaturase-like [Homo sapiens] 14 BAD96861 62897843 sterol-C5-desaturase (ERG3 delta-5-desaturase homolog, fungal)- like variant [Homo sapiens] 15 BAD96406 62896931 sterol-C5-desaturase (ERG3 delta-5-desaturase homolog, fungal)- like variant [Homo sapiens] 16 AAH50427 30046554 Sterol-C5-desaturase (ERG3 delta-5-desaturase homolog, fungal)- like [Homo sapiens] 17 AAH12333 15147389 Sterol-C5-desaturase (ERG3 delta-5-desaturase homolog, fungal)- like [Homo sapiens] 18 AAF00544 6003685 delta7-sterol-C5-desaturase [Homo sapiens] 19 BAB68218 15637108 sterol C5 desaturase [Homo sapiens] 20 O75845 6174975 Lathosterol oxidase (Lathosterol 5-desaturase) (Delta-7-sterol 5- desaturase) (C-5 sterol desatur 21 BAA33729 3721882 sterol-C5-desaturase [Homo sapiens] 22 XP_001 . . . 1.09E+08 PREDICTED: sterol-C5-desaturase (ERG3 delta-5-desaturase homolog, fungal)-like isoform 2 [Macaca 23 AAH78055 50416254 Sc5d-prov protein [Xenopus laevis] 24 NP_001 . . . 52219112 sterol-C5-desaturase (fungal ERG3, delta-5-desaturase) homolog [Danio rerio] 25 AAH81395 51859008 Sterol-C5-desaturase (fungal ERG3, delta-5-desaturase) homolog (S. cerevisae) [Danio rerio] 26 BAA18970 1906796 fungal sterol-C5-desaturase homolog [Homo sapiens] 27 XP_001 . . . 1.09E+08 PREDICTED: sterol-C5-desaturase (ERG3 delta-5-desaturase homolog, fungal)-like isoform 1 [Macaca 28 XP_793331 72159211 PREDICTED: similar to sterol-C5-desaturase (fungal ERG3, delta- 5-desaturase) homolog [Strongyloc 29 NP_001 . . . 78369462 hypothetical protein LOC525154 [Bos taurus] 30 AAI03339 74353920 Similar to sterol-C5-desaturase-like [Bos taurus] 31 CAF90498 47207599 unnamed protein product [Tetraodon nigroviridis]

TABLE 61 Examples of ERG27 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 Q12452 60392263 3-keto-steroid reductase 2 CAA97664 1360483 unnamed protein product [Saccharomyces cerevisiae] 3 AAB67544 1256850 Ylr100wp [Saccharomyces cerevisiae] 4 Q6FIV3 62286920 3-keto-steroid reductase 5 P_449841 50294860 hypothetical protein CAGL0M11506g [Candida glabrata CBS138] 6 CAG62821 49529155 unnamed protein product [Candida glabrata CBS138] 7 Q6CJC2 62286919 3-keto-steroid reductase 8 P_455967 50311871 unnamed protein product [Kluyveromyces lactis] 9 CAG98675 49645103 unnamed protein product [Kluyveromyces lactis NRRL Y-1140] 10 Q74ZZ0 62286926 3-keto-steroid reductase 11 P_986733 45201163 AGR068Wp [Eremothecium gossypii] 12 AAS54557 44985946 AGR068Wp [Ashbya gossypii ATCC 10895] 13 Q6BNP0 62286917 3-keto-steroid reductase 14 P_460180 50423199 hypothetical protein DEHA0E21263g [Debaryomyces hansenii CBS 15 CAG88453 49655848 unnamed protein product [Debaryomyces hansenii CBS767] 16 Q874K0 62286935 3-keto-steroid reductase 17 AAN28009 29465858 3-keto reductase [Candida albicans] 18 P_717865 68476112 3-keto sterol reductase [Candida albicans SC5314] 19 P_717931 68475981 3-keto sterol reductase [Candida albicans SC5314] 20 EAK98982 46439667 hypothetical protein CaO19.3240 [Candida albicans SC5314] 21 EAK98915 46439599 hypothetical protein CaO19.10750 [Candida albicans SC5314] 22 Q6CE88 62286918 3-keto-steroid reductase 23 P_501024 50547109 hypothetical protein [Yarrowia lipolytica] 24 CAG83277 49646890 unnamed protein product [Yarrowia lipolytica CLIB122] 25 P_595440 19112232 3-keto sterol reductase [Schizosaccharomyces pombe 972h-] 26 CAA21246 3738145 SPBC1709.07 [Schizosaccharomyces pombe] 27 BAA13878 1749642 unnamed protein product [Schizosaccharomyces pombe] 28 P_751680 70993666 3-ketosteroid reductase [Aspergillus fumigatus Af293] 29 EAL89642 66849314 3-ketosteroid reductase [Aspergillus fumigatus Af293] 30 P_958799 85091228 hypothetical protein [Neurospora crassa OR74A] 31 EAA29563 28920185 hypothetical protein [Neurospora crassa] 32 BAE58171 83768032 unnamed protein product [Aspergillus oryzae] 33 EAQ85086 88177618 hypothetical protein CHGG_09100 [Chaetomium globosum CBS 148 34 EAT86025 111064905 hypothetical protein SNOG_06194 [Phaeosphaeria nodorum SN15] 35 P_663189 67538830 hypothetical protein AN5585.2 [Aspergillus nidulans FGSC A4] 36 EAA62228 40743038 hypothetical protein AN5585.2 [Aspergillus nidulans FGSC A4] 37 EAS36221 90306590 hypothetical protein CIMG_01575 [Coccidioides immitis RS] 38 AAO64345 29150686 3-ketosteroid reductase [Botryotinia fuckeliana] 39 P_796770 72131681 PREDICTED: similar to hydroxysteroid (17-beta) dehydrogenase 40 P_793145 72086766 PREDICTED: similar to hydroxysteroid (17-beta) dehydrogenase 41 P_570518 58266724 3-keto sterol reductase [Cryptococcus neoformans var. neofor 42 AAW43211 57226751 3-keto sterol reductase, putative [Cryptococcus neoformans v 43 EAL21279 50258592 hypothetical protein CNBD3330 [Cryptococcus neoformans var. 44 AAI00220 71681260 Unknown (protein for MGC: 114978) [Xenopus laevis] 45 BAE06843 70720928 hypothetical protein [Epichloe festucae] 46 P_001 . . . 62859081 hydroxysteroid (17-beta) dehydrogenase 7 [Xenopus tropicalis 47 P_851698 74006237 PREDICTED: similar to hydroxysteroid (17-beta) dehydrogenase 48 CAC88119 18077845 17 beta-hydroxysteroid dehydrogenase type 7 [Mus musculus] 49 AAH11464 15079256 Hsd17b7 protein [Mus musculus] 50 P_034606 87162470 hydroxysteroid (17-beta) dehydrogenase 7 [Mus musculus] 51 BAC25918 26390565 unnamed protein product [Mus musculus] 52 BAC34124 26340924 unnamed protein product [Mus musculus] 53 O88736 8134403 3-keto-steroid reductase (Estradiol 17-beta-dehydrogenase 7) 54 CAA75742 33199711 7-beta-hydroxysteroid dehydrogenase type 7 [Mus musculus] 55 P_695820 68440091 PREDICTED: similar to Hydroxysteroid (17-beta) dehydrogenase 56 P_058931 8393576 hydroxysteroid (17-beta) dehydrogenase 7 [Rattus norvegicus] 57 Q62904 8134405 3-keto-steroid reductase (Estradiol 17-beta-dehydrogenase 7) 58 AAC52623 1397235 ovarian-specific protein 59 BAC37061 26346825 unnamed protein product [Mus musculus] 60 AAM21211 203851961 7-beta-hydroxysteroid dehydrogenase type 7 [Mus musculus] 61 P_363377 39951321 hypothetical protein MG01303.4 [Magnaporthe grisea 70-15] 62 P_581467 76611932 PREDICTED: similar to hydroxysteroid (17-beta) dehydrogenase 63 CAC88111 18077843 17 beta hydroxysteroid dehydrogenase [Homo sapiens] 64 AAX42032 61361334 hydroxysteroid (17-beta) dehydrogenase 7 [synthetic construc 65 AAX42031 61361326 hydroxysteroid (17-beta) dehydrogenase 7 [synthetic construc 66 CAI13457 55959848 hydroxysteroid (17-beta) dehydrogenase 7 [Homo sapiens] 67 CAI15947 55959770 hydroxysteroid (17-beta) dehydrogenase 7 [Homo sapiens] 68 AAH65246 40807207 Hydroxysteroid (17-beta) dehydrogenase 7 [Homo sapiens] 69 AAQ89321 37183042 HSD17B7 [Homo sapiens] 70 AAP35738 30582989 hydroxysteroid (17-beta) dehydrogenase 7 [Homo sapiens] 71 AAH07068 13937918 Hydroxysteroid (17-beta) dehydrogenase 7 [Homo sapiens] 72 CAC20418 12053691 17 beta-hydroxysteroid dehydrogenase type 7 [Homo sapiens] 73 P56937 8134404 3-keto-steroid reductase (Estradiol 17-beta-dehydrogenase 7) 74 P_057455 7705421 hydroxysteroid (17-beta) dehydrogenase 7 [Homo sapiens] 75 AAF09266 6721095 17 beta-hydroxysteroid dehydrogenase type VII [Homo sapiens] 76 ABB72853 82400271 17-beta hydroxysteroid dehydrogenase 7 [Macaca fascicularis] 77 AAX29487 60653587 hydroxysteroid (17-beta) dehydrogenase 7 [synthetic construc 78 AAP36180 30583863 Homo sapiens hydroxysteroid (17-beta) dehydrogenase 7 [synth 79 AAF14537 6502988 17 beta-hydroxysteroid dehydrogenase type 7 [Homo sapiens] 80 AAK20950 13383374 17-beta-hydroxysteroid dehydrogenase type 7 [Oryctolagus cun 81 AAP97275 33150794 ovarian-specific protein; OSP [Homo sapiens] 82 P_422210 50750970 PREDICTED: similar to Estradiol 17 beta-dehydrogenase 7 (17-

TABLE 62 Examples of ERG26 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAT93045 51013503 YGL001C [Saccharomyces cerevisiae] 2 P53199 1723793 Sterol-4-alpha-carboxylate 3-dehydrogenase, decarboxylating 3 CAA96701 1322447 unnamed protein product [Saccharomyces cerevisiae] 4 P_446390 50287923 unnamed protein product [Candida glabrata] 5 CAG59317 49525698 unnamed protein product [Candida glabrata CBS138] 6 P_453025 50306127 unnamed protein product [Kluyveromyces lactis] 7 CAH01876 49642158 unnamed protein product [Kluyveromyces lactis NRRL Y-1140] 8 CAC38016 13940379 putative C-3 sterol dehydrogenase [Zygosaccharomyces rouxii] 9 P_985548 45198519 AFR001 Wp [Eremothecium gossypii] 10 AAS53372 44984470 AFR001Wp [Ashbya gossypii ATCC 10895] 11 P_458178 50419307 hypothetical protein DEHA0C12463g [Debaryomyces hansenii CBS767] 12 CAG86252 49653844 unnamed protein product [Debaryomyces hansenii CBS767] 13 P_715564 68481045 C-3 sterol dehydrogenase/C-4 decarboxylase [Candida albicans SC53 14 P_715620 68480934 C-3 sterol dehydrogenase/C-4 decarboxylase [Candida albicans SC53 15 EAK96601 46437251 hypothetical protein CaO19.10427 [Candida albicans SC5314] 16 EAK96542 46437191 hypothetical protein CaO19.2909 [Candida albicans SC5314] 17 AAK69617 14582743 C-3 sterol dehydrogenase/C-4 decarboxylase [Candida albicans] 18 P_502124 50549307 hypothetical protein [Yarrowia lipolytica] 19 CAG82444 49647991 unnamed protein product [Yarrowia lipolytica CLIB122] 20 P_596741 19113533 hypothetical protein SPBC3F6.02c [Schizosaccharomyces pombe 972h- 21 CAA17691 2924499 SPBC3F6.02c [Schizosaccharomyces pombe] 22 AAQ88129 37039511 C-3 sterol dehydrogenase [Cryptococcus neoformans var. grubii H99 23 P_381379 46108642 hypothetical protein FG01203.1 [Gibberella zeae PH-1] 24 EAA67667 42544824 hypothetical protein FG01203.1 [Gibberella zeae PH-1] 25 EAQ90884 88183416 conserved hypothetical protein [Chaetomium globosum CBS 148.51] 26 P_640712 66813066 3 beta-hydroxysteroid dehydrogenase [Dictypstelium discoideum AX4] 27 EAL66725 60468723 3 beta-hydroxysteroid dehydrogenase [Dictypstelium discoideum AX4] 28 EAT77489 111056369 hypothetical protein SNOG_15264 [Phaeosphaeria nodorum SN15] 29 P_965466 85118539 hypothetical protein [Neurospora crassa OR74A] 30 CAE76518 38567226 related to C-3 sterol dehydrogenase (C-4 decarboxylase) [Neurospo 31 EAA36230 28927275 hypothetical protein [Neurospora crassa] 32 P_420279 50745878 PREDICTED: similar to NAD(P) dependent steroid dehydrogenase-like 33 P_001 . . . 78369400 hypothetical protein LOC616694 [Bos taurus] 34 AAI03390 73586574 Similar to NAD(P) dependent steroid dehydrogenase-like [Bos tauru 35 P_001 . . . 62859757 hypothetical protein LOC550044 [Xenopus tropicalis] 36 AAH88699 56788986 LOC496236 protein [Xenopus laevis] 37 P_538199 74008671 PREDICTED: similar to NAD(P) dependent steroid dehydrogenase-like 38 P_853073 74008673 PREDICTED: similar to NAD(P) dependent steroid dehydrogenase-like 39 AAH52834 31127258 NAD(P) dependent steroid dehydrogenase-like [Mus musculus] 40 P_001 . . . 109132673 PREDICTED: similar to NAD(P) dependent steroid dehydrogenase-like 41 P_001 . . . 109132671 PREDICTED: similar to NAD(P) dependent steroid dehydrogenase-like 42 P_001 . . . 57164113 NAD(P) dependent steroid dehydrogenase-like [Rattus norvegicus] 43 AAH87626 56388600 NAD(P) dependent steroid dehydrogenase-like [Rattus norvegicus] 44 BAE24522 74187675 unnamed protein product [Mus musculus] 45 P_035071 31982437 NAD(P) dependent steroid dehydrogenase-like [Mus musculus] 46 Q9R1J0 8473695 Sterol-4-alpha-carboxylate 3-dehydrogenase, decarboxylating 47 CAA15948 7619723 NAD(P)H steroid dehydrogenase [Mus musculus] 48 AAD38448 5052206 putative NAD(P)H steroid dehydrogenase [Mus musculus] 49 AAH19945 18043286 NAD(P) dependent steroid dehydrogenase-like [Mus musculus] 50 AAH07816 14043700 NAD(P) dependent steroid dehydrogenase-like [Homo sapiens] 51 AAH00245 12652969 NAD(P) dependent steroid dehydrogenase-like [Homo sapiens] 52 Q15738 8488997 Sterol-4-alpha-carboxylate 3-dehydrogenase, decarboxylating (H105 53 P_057006 8393516 NAD(P) dependent steroid dehydrogenase-like [Homo sapiens] 54 AAC50558 4457237 H105e3 [Homo sapiens] 55 CAF99107 47222951 unnamed protein product [Tetraodon nigroviridis] 56 P_756075 71002788 C-3 sterol dehydrogenase/C-4 decarboxylase [Aspergillus fumigatus 57 EAL94037 66853713 C-3 sterol dehydrogenase/C-4 decarboxylase [Aspergillus fumigatus 58 P_567299 58259773 C-3 sterol dehydrogenase (C-4 sterol decarboxylase) [Cryptococcus 59 AAW45782 57229349 C-3 sterol dehydrogenase (C-4 sterol decarboxylase), putative [Cr 60 EAL18383 50255650 hypothetical protein CNBJ3060 [Cryptococcus neoformans var. neofo 61 P_001 . . . 62955325 hypothetical protein LOC550369 [Danio rerio] 62 AAH93332 62204675 Zgc:112474 [Danio rerio] 63 EAS37011 90307380 hypothetical protein CIMG_02365 [Coccidioides immitis RS] 64 P_755836 71002310 C-3 sterol dehydrogenase/C-4 decarboxylase [Aspergillus fumigatus 65 EAL93798 66853474 C-3 sterol dehydrogenase/C-4 decarboxylase [Aspergillus fumigatus 66 CAH90815 55728130 hypothetical protein [Pongo pygmaeus] 67 P_760274 71020087 hypothetical protein UM04127.1 [Ustilago maydis 521] 68 EAK85155 46099922 hypothetical protein UM04127.1 [Ustilago maydis 521] 69 P_680844 67901176 hypothetical protein AN7575.2 [Aspergillus nidulans FGSC A4] 70 EAA62155 40742965 hypothetical protein AN7575.2 [Aspergillus nidulans FGSC A4] 71 AAZ14936 70906332 putative sterol dehydrogenase [Coprinellus disseminatus]

TABLE 63 Examples of ERG25 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAS56153 45269545 YGR060W [Saccharomyces cerevisiae] 2 XP_448420 50291975 unnamed protein product [Candida glabrata] 3 XP_451890 50303885 unnamed protein product [Kluyveromyces lactis] 4 NP_986119 45199090 AFR572Wp [Eremothecium gossypii] 5 XP_456861 50409318 hypothetical protein DEHA0A12661g [Debaryomyces hansenii CBS767] 6 XP_713420 68485318 C-4 sterol methyl oxidase [Candida albicans SC5314] 7 XP_461658 50426123 hypothetical protein DEHA0G03124g [Debaryomyces hansenii CBS767] 8 XP_505281 50555746 hypothetical protein [Yarrowia lipolytica] 9 XP_722703 68466442 putative C-4 sterol methyl oxidase [Candida albicans SC5314] 10 XP_722849 68466149 putative C-4 sterol methyl oxidase [Candida albicans SC5314] 11 ABF84061 107785180 C-4 sterol methyl oxidase [Chaetomium globosum] 12 Q9UUH4 28558117 C-4 methylsterol oxidase (Methylsterol monooxygenase) 13 XP_962240 85106720 hypothetical protein [Neurospora crassa OR74A] 14 EAT84680 111063560 hypothetical protein SNOG_08404 [Phaeosphaeria nodorum SN15] 15 XP_746668 70982279 hypothetical protein Afu4g04820 [Aspergillus fumigatus Af293] 16 XP_390006 46136629 hypothetical protein FG09830.1 [Gibberella zeae PH-1] 17 BAE65381 83775259 unnamed protein product [Aspergillus oryzae] 18 XP_360827 39942580 hypothetical protein MG03370.4 [Magnaporthe grisea 70-15] 19 ABA70590 77377695 hypothetical protein [Penicillium chrysogenum] 20 XP_369331 39975881 hypothetical protein MG06133.4 [Magnaporthe grisea 70-15] 21 EAS29762 90300131 hypothetical protein CIMG_08508 [Coccidioides immitis RS] 22 XP_762027 71023595 hypothetical protein UM05880.1 [Ustilago maydis 521] 23 XP_569526 58264740 C-4 methyl sterol oxidase [Cryptococcus neoformans var. neoformans JEC21] 24 EAQ88125 88180657 hypothetical protein CHGG_04744 [Chaetomium globosum CBS 148.51] 25 BAE66491 83776372 unnamed protein product [Aspergillus oryzae] 26 XP_746962 70982869 c-4 methyl sterol oxidase [Aspergillus fumigatus Af293] 27 XP_682176 67903840 hypothetical protein AN8907.2 [Aspergillus nidulans FGSC A4] 28 XP_664577 67541619 hypothetical protein AN6973.2 [Aspergillus nidulans FGSC A4] 29 XP_646277 66825845 hypothetical protein DDBDRAFT_0190553 [Dictyostelium discoideum AX4] 30 NP_998917 47523336 sterol-C4-methyl oxidase-like protein [Sus scrofa] 31 AAI07880 79160074 Sterol-C4-methyl oxidase-like [Homo sapiens] 32 BAE01921 67971158 unnamed protein product [Macaca fascicularis] 33 XP_001 . . . 109076099 PREDICTED: sterol-C4-methyl oxidase-like isoform 3 [Macaca mulatta] 34 XP_580615 61871492 PREDICTED: similar to C-4 methylsterol oxidase (Methylsterol monooxygenase) isoform 1 [Bos taurus] 35 CAC24471 12311696 hypothetical protein [Candida albicans] 36 BAE30143 74185334 unnamed protein product [Mus musculus] 37 XP_532714 57096891 PREDICTED: similar to C-4 methylsterol oxidase (Methylsterol monooxygenase) isoform 1 [Canis familiaris] 38 CAF92153 47211304 unnamed protein product [Tetraodon nigroviridis] 39 AAH63155 38649308 Sterol-C4-methyl oxidase-like [Rattus norvegicus] 40 NP_001 . . . 57530155 sterol-C4-methyl oxidase-like [Gallus gallus] 41 NP_998518 47087009 sterol-C4-methyl oxidase-like [Danio rerio] 42 NP_563789 18390767 SMO2-2; C-4 methylsterol oxidase [Arabidopsis thaliana] 43 AAI21457 111309060 Unknown (protein for MGC: 146461) [Xenopus tropicalis] 44 AAM64821 21592871 putative C-4 sterol methyl oxidase [Arabidopsis thaliana] 45 AAF79571 8778563 F22G5.23 [Arabidopsis thaliana] 46 AAO13795 27448145 putative sterol 4-alpha-methyl-oxidase [Gossypium arboreum] 47 AAO48604 28864263 ERG25 [Clavispora lusitaniae] 48 AAQ83692 34978966 C-4 sterol methyl oxidase 2 [Nicotiana benthamiana] 49 ABD97153 90658509 C-4 methyl sterol oxidase [Cryptococcus neoformans var. neoformans] 50 ABD97146 90658495 C-4 methyl sterol oxidase [Cryptococcus neoformans var. grubii] 51 ABD97147 90658497 C-4 methyl sterol oxidase [Cryptococcus neoformans var. grubii] 52 ABD97134 90658471 C-4 methyl sterol oxidase [Cryptococcus neoformans var. neoformans] 53 AAL82576 27447203 putative sterol 4-alpha-methyl-oxidase [Zea mays] 54 NP_850133 30684225 SMO2-1; C-4 methylsterol oxidase [Arabidopsis thaliana] 55 NP_973559 42570971 SMO2-1; C-4 methylsterol oxidase [Arabidopsis thaliana] 56 AAL32287 16973432 sterol 4-alpha-methyl-oxidase [Arabidopsis thaliana] 57 ABE83226 92875070 Sterol desaturase [Medicago truncatula] 58 BAF28920 113645779 Os11g0707600 [Oryza sativa (japonica cultivar-group)] 59 BAF20592 113610214 Os07g0101500 [Oryza sativa (japonica cultivar-group)] 60 XP_787442 72077919 PREDICTED: similar to C-4 methylsterol oxidase (Methylsterol monooxygenase) [Strongylocentrotus purpuratus] 61 AAM64359 21592408 putative C-4 sterol methyl oxidase [Arabidopsis thaliana] 62 NP_973777 42571373 SMO2-2; C-4 methylsterol oxidase [Arabidopsis thaliana] 63 NP_001 . . . 62865628 sterol-C4-methyl oxidase-like isoform 2 [Homo sapiens] 64 AAX95419 62733302 4-alpha-methyl-sterol C4-methyl-oxidase [Oryza sativa (japonica cultivar-group)] 65 XP_883875 76638226 PREDICTED: similar to C-4 methylsterol oxidase (Methylsterol monooxygenase) isoform 3 [Bos taurus] 66 ABE87434 92884635 Sterol desaturase [Medicago truncatula] 67 XP_641553 66814748 hypothetical protein DDBDRAFT_0205936 [Dictyostelium discoideum AX4] 68 AAM65428 21593461 putative C-4 sterol methyl oxidase [Arabidopsis thaliana] 69 NP_567669 18416002 SMO1-3 (STEROL 4-alpha METHYL OXIDASE); catalytic [Arabidopsis thaliana] 70 AAM64961 21593012 putative C-4 sterol methyl oxidase [Arabidopsis thaliana] 71 AAN18115 23308291 At4g12110/F16J13_180 [Arabidopsis thaliana] 72 AAK61361 27446631 putative sterol 4-alpha-methyl-oxidase [Arabidopsis thaliana] 73 BAF01201 110738551 hypothetical protein [Arabidopsis thaliana] 74 AAQ13424 33337546 sterol-4-methyl-oxidase [Arabidopsis thaliana] 75 BAF27120 113639815 Os10g0545200 [Oryza sativa (japonica cultivar-group)] 76 CAB78254 7267912 putative C-4 sterol methyl oxidase [Arabidopsis thaliana] 77 AAQ94118 37591406 sterol-4-alpha methyl oxidase [Arabidopsis thaliana] 78 AAQ83691 34978964 C-4 sterol methyl oxidase 1 [Nicotiana benthamiana] 79 ABD65536 89266491 sterol-C4-methyl oxidase-like [Ictalurus punctatus] 80 BAC57961 28804511 putative C-4 sterol methyl oxidase [Aster tripolium]

TABLE 64 Examples of ERG24 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAB30203 545906 sterol delta 14 reductase; Erg24p [Saccharomyces cerevisiae] 2 XP_447380 50289897 hypothetical protein CAGL0I02970g [Candida glabrata CBS138] 3 XP_452744 50305569 unnamed protein product [Kluyveromyces lactis] 4 NP_985893 45198864 AFR346Wp [Eremothecium gossypii] 5 XP_710670 68491033 sterol C-14 reductase [Candida albicans SC5314] 6 XP_458371 50419687 hypothetical protein DEHA0C16984g [Debaryomyces hansenii CBS767] 7 XP_501260 50547581 hypothetical protein [Yarrowia lipolytica] 8 XP_566548 58258271 C-14 sterol reductase [Cryptococcus neoformans var. neoformans JEC21] 9 NP_596767 19113559 hypothetical protein SPBC16G5.18 [Schizosaccharomyces pombe 972h-] 10 EAL23454 50260804 hypothetical protein CNBA1040 [Cryptococcus neoformans var. neoformans B-3501A] 11 XP_759786 71019111 hypothetical protein UM03639.1 [Ustilago maydis 521] 12 XP_386782 46124457 ER24_FUSSO Delta(14)-sterol reductase (C-14 sterol reductase) (Sterol C14-reductase) [Gibberella zeae PH-1] 13 P_78575 6015106 Delta(14)-sterol reductase (C-14 sterol reductase) (Sterol C14- reductase) 14 Q01447 3929349 Delta(14)-sterol reductase (C-14 sterol reductase) (Sterol C14- reductase)

TABLE 65 Examples of ERG11 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAA34546 171354 lanosterol 14 -demethylase cyto chrome P450 2 AAA34547 171356 lanosterol 14 -demethylase cyto chrome P450 3 AAY59417 66967944 14 alpha-demethylase [Candida glabrata] 4 XP_454109 50308217 unnamed protein product [Kluyveromyces lactis] 5 Q759W0 51701328 Cytochrome P450 51 (CYPLI) (P450-LIA1) (Sterol 14-alpha demethylase) (Lanosterol 14-alpha demethylase) (P450-14DM) 6 AAB32679 912807 cytochrome P-450 lanosterol-alpha-demethylase; P450-L1A1 [Candida glabrata] 7 AAX39316 61189834 lanosterol 14 alpha demethylase [Candida tropicalis] 8 AAF00598 6006769 cytochrome P450 lanosterol 14-alpha demethylase [Candida albicans] 9 XP_716761 68478280 lanosterol 14-alpha-demethylase [Candida albicans SC5314] 10 XP_460143 50423125 hypothetical protein DEHA0E20383g [Debaryomyces hansenii CBS767] 11 ABC41921 83728463 cytochrome P450 lanosterol 14 alpha demethylase [Candida albicans] 12 AAX39315 61189832 lanosterol 14 alpha demethylase [Candida tropicalis] 13 O4CK51 65689 lanosterol 14 alpha-demethylase (EC 1.14.14.-) cytochrome P450 51- yeast (Candida albicans) 14 XP_716822 68478161 lanosterol 14-alpha-demethylase [Candida albicans SC5314] 15 AAF00603 6006774 cytochrome P450 lanosterol 14-alpha demethylase [Candida albicans] 16 AAF00600 6006771 cytochrome P450 lanosterol 14-alpha demethylase [Candida albicans] 17 BAB03401 9558446 CYP51 variant 3 [Candida albicans] 18 AAK57519 14280372 lanosterol 14-alpha demethylase Erg11p [Candida dubliniensis] 19 AAF00602 6006773 cytochrome P450 lanosterol 14-alpha demethylase [Candida albicans] 20 AAF00601 6006772 cytochrome P450 lanosterol 14-alpha demethylase [Candida albicans] 21 BAB03400 9558444 CYP51 variant 2 [Candida albicans] 22 AAO83898 29378393 cytochrome P-450 lanosterol-alpha-demethylase [Issatchenkia orientalis] 23 AAW50593 57341382 Erg11p [Candida albicans] 24 AAW50592 57341380 Erg11p [Candida albicans] 25 XP_500518 50545962 hypothetical protein [Yarrowia lipolytica] 26 BAC16517 23263326 cytochrome P-450 lanosterol 14 alpha-demethylase [Candida albicans] 27 BAC16520 23263332 cytochrome P-450 lanosterol 14 alpha-demethylase [Candida albicans] 28 BAC16518 23263328 cytochrome P-450 lanosterol 14 alpha-demethylase [Candida albicans] 29 BAC16519 23263330 cytochrome P-450 lanosterol 14 alpha-demethylase [Candida albicans] 30 Q02315 2493385 Cytochrome P450 51 (CYPLI) (P450-LIA1) (Sterol 14-alpha demethylase) (Lanosterol 14-alpha demethylase) (P450-14DM) 31 AAO38776 28395121 lanosterol 14 alpha demethylase; ERG11 [Pneumocystis carinii] 32 ABG91757 110816094 sterol 14 alpha-demethylase [Pneumocystis carinii] 33 BAE60239 83770104 unnamed protein product [Aspergillus oryzae] 34 XP_749134 70987441 14-alpha sterol demethylase Cyp51B [Aspergillus fumigatus Af293] 35 XP_659505 67522889 hypothetical protein AN1901.2 [Aspergillus nidulans FGSC A4] 36 XP_681552 67902592 hypothetical protein AN8283.2 [Aspergillus nidulans FGSC A4] 37 AAK73659 14861413 14-alpha sterol demethylase [Aspergillus fumigatus] 38 XP_752137 70994720 14-alpha sterol demethylase Cyp51A [Aspergillus fumigatus Af293] 39 BAE57417 83767278 unnamed protein product [Aspergillus oryzae] 40 AAF79204 8778097 cytochrome P450 sterol 14 alpha-demethylase [Emericella nidulans] 41 CAE18091 33300264 eburicol 14 alpha demethylase [Blumeria graminis] 42 EAS28723 90299092 cytochrome P450 51 [Coccidioides immitis RS] 43 CAE18094 33300270 eburicol 14 alpha demethylase [Blumeria graminis] 44 CAC85624 33300256 eburicol 14 alpha demethylase [Blumeria graminis f. sp. tritici] 45 CAE18095 33300272 eburicol 14 alpha demethylase [Blumeria graminis] 46 CAE17515 33284834 eburicol 14 alpha demethylase [Blumeria graminis f. sp. tritici] 47 AAU43734 52352491 eburicol 14 alpha-demethylase [Mycosphaerella graminicola] 48 EAS35219 90305588 cytochrome P450 51 [Coccidioides immitis RS] 49 CAE18102 33300286 eburicol 14 alpha demethylase [Blumeria graminis] 50 CAE18103 33300288 eburicol 14 alpha demethylase [Blumeria graminis] 51 CAE18105 33300292 eburicol 14 alpha demethylase [Blumeria graminis] 52 NP_592990 19113902 hypothetical protein SPAC13A11.02c [Schizosaccharomyces pombe 972h-] 53 CAE18100 33300282 eburicol 14 alpha demethylase [Blumeria graminis] 54 AAP79601 32330152 eburicol 14 alpha-demethylase [Mycosphaerella graminicola] 55 AAC97606 4049645 eburicol 14 alpha demethylase; CYP51; cytochrome P450 sterol 14- demethylase [Blumeria graminis f. sp. hordei] 56 CAE18099 33300280 eburicol 14 alpha demethylase [Blumeria graminis] 57 BAE63554 83773427 unnamed protein product [Aspergillus oryzae] 58 XP_381176 46108236 hypothetical protein FG01000.1 [Gibberella zeae PH-1] 59 AAF85983 9230788 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 60 AAP81934 32441730 eburicol 14 alpha-demethylase [Mycosphaerella graminicola] 61 XP_362183 39945292 hypothetical protein MG04628.4 [Magnaporthe grisea 70-15] 62 AAP12370 30314338 lanosterol 14 alpha-demethylase; 14 alpha lanosterol demethylase [Cryptococcus neoformans var. grubii] 63 AAF74756 8347731 eburicol 14-alpha demethylase [Mycosphaerella graminicola] 64 AAF76464 8570176 14 alpha-demethylase [Venturia inaequalis] 65 XP_566464 58258103 sterol 14-demethylase [Cryptococcus neoformans var. neoformans JEC21] 66 EAQ93417 88185949 conserved hypothetical protein [Chaetomium globosum CBS 148.51] 67 AAL79180 18766932 eburicol 14 alpha-demethylase [Monilinia fructicola] 68 AAU01157 51341094 lanosterol 14-alpha-demethylase [Coccidioides posadasii] 69 CAD27793 19572745 cytochrome P-450 14DM [Penicillium digitatum] 70 XP_384268 46116500 hypothetical protein FG04092.1 [Gibberella zeae PH-1] 71 AAG18433 10505323 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 72 AAK26391 13445823 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 73 AAG18431 10505318 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 74 AAK26386 13445813 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 75 AAD55135 5881952 eburicol 14-alpha demethylase; cytochrome P450 sterol 14-alpha demethylase; Erg11 [Uncinula necator] 76 AAK26389 13445819 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 77 CAC85409 15864613 euburicol 14 alpha-demethylase [Venturia nashicola] 78 BAB03659 9664027 cytochrome P-450 14DM [Penicillium digitatum] 79 AAG18435 10505329 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 80 AAG18434 10505326 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 81 AAU01158 51341096 lanosterol 14-alpha-demethylase [Ajellomyces capsulatus] 82 AAK26387 13445815 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 83 AAG18432 10505321 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 84 AAK26388 13445817 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 85 XP_391200 46139019 hypothetical protein FG11024.1 [Gibberella zeae PH-1] 86 AAK26390 13445821 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 87 O14442 14916976 Cytochrome P450 51 (CYPLI) (P450-LIA1) (Sterol 14-alpha demethylase) (Eburicol 14-alpha-demethylase) (P450-14DM) 88 AAG18438 10505336 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 89 XP_759809 71019157 sterol 14-alpha demethylase [Ustilago maydis 521] 90 AAF18468 6581118 eburicol 14 alpha-demethylase [Tapesia acuformis] 91 AAG18436 10505331 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 92 AAC49801 5733843 eburicol 14-alpha demethylase [Uncinula necator] 93 ABE01107 908232111 4 alpha demethylase [Blumeriella jaapii] 94 AAG18437 10505334 eburicol 14 alpha-demethylase [Botryotinia fuckeliana] 95 CAD27792 19572743 cytochrome P-450 14DM [Penicillium digitatum] 96 XP_964049 85111674 hypothetical protein [Neurospora crassa OR74A] 97 AAF18469 11178694 eburicol 14 alpha-demethylase [Tapesia yallundae] 98 Q12664 2493386 Cytochrome P450 51 (CYPLI) (P450-LIA1) (Sterol 14-alpha demethylase) (Eburicol 14-alpha-demethylase) (P450-14DM) 99 AAG44832 12006321 eburicol 14-alpha demethylase [Tapesia yallundae] 100 AAG44831 12006319 eburicol 14-alpha demethylase [Tapesia yallundae] 101 AAN28927 23505745 sterol 14 alpha-demethylase ERG11 [Leptosphaeria maculans] 102 XP_361987 39944900 hypothetical protein MG04432.4 [Magnaporthe grisea 70-15] 103 EAT88907 111067787 hypothetical protein SNOG_03702 [Phaeosphaeria nodorum SN15] 104 AAU01160 51341100 lanosterol 14-alpha-demethylase [Phanerochaete chrysosporium] 105 AAC23550 3220152 cytochrome P450 L1A1 demethylase [Pichia anomala] 106 AAU01159 51341098 lanosterol 14-alpha-demethylase [Coprinopsis cinerea] 107 Q9UVC3 12229768 Cytochrome P450 51 (CYPLI) (P450-LIA1) (Sterol 14-alpha demethylase) (Lanosterol 14-alpha demethylase) (P450-14DM) 108 AAQ88128 37039509 cytochrome P450 lanosterol 14a-demethylase [Cryptococcus neoformans var. grubii H99] 109 NP_064394 71061451 cytochrome P450, family 51 [Mus musculus] 110 EAL24154 51094909 cytochrome P450, family 51, subfamily A, polypeptide 1 [Homo sapiens] 111 Q16850 3915660 Cytochrome P450 51A1 (CYPLI) (P45011) (Sterol 14-alpha demethylase) (Lanosterol 14-alpha demethylase) (LDM) (P450- 14DM) (P45014DM) 112 NP_999597 47523914 cytochrome P450 51 [Sus scrofa] 113 BAC27231 26326975 unnamed protein product [Mus musculus] 114 Q5RE72 83287777 Cytochrome P450 51A1 (CYPLI) (P45011) (Sterol 14-alpha demethylase) (Lanosterol 14-alpha demethylase) (LDM) (P450- 14DM) (P45014DM) 115 Q4PJW3 7507506614 Cytochrome P450 51A1 (CYPLI) (P45011) (Sterol 14-alpha demethylase) (Lanosterol 14-alpha demethylase) (LDM) (P450- DM) (P45014DM) 116 AAF74562 8347236 lanosterol 14-alpha-demethylase [Mus musculus] 117 AAC50951 1698484 lanosterol 14-alpha demethylase [Homo sapiens] 118 XP_532457 57095976 PREDICTED: similar to Cytochrome P450 51A1 (CYPLI) (P450LI) (Sterol 14-alpha demethylase) (Lanosterol 14-alpha demethylase) (LDM) (P450-14DM) (P45014DM) isoform 1 [Canis familiaris] 119 AAF73986 8215679 lanosterol 14-alpha-demethylase [Mus musculus] 120 AAH81210 51703589 MGC84806 protein [Xenopus laevis] 121 AAA87074 699396 lanosterol 14-alpha-demethylase 122 XP_418650 50732469 PREDICTED: similar to lanosterol 14-demethylase [Gallus gallus] 123 BAE01508 67970330 unnamed protein product [Macaca fascicularis] 124 AAR89624 40748287 14-alpha demethylase [Fundulus heteroclitus] 125 NP_001 . . . 50053648 cytochrome P450, family 51 [Strongylocentrotus purpuratus] 126 AAR89625 40748289 14-alpha demethylase [Danio rerio] 127 NP_001 . . . 99028936 cytochrome P450, family 51 [Danio rerio] 128 CAJ81841 89268132 cytochrome P450, family 51, subfamily A, polypeptide 1 [Xenopus tropicalis] 129 AAR89626 40748291 14-alpha demethylase [Abudefduf saxatilis]

TABLE 66 Examples of ERG4 polypeptides. Row ACCESSION GI PROTEIN DESCRIPTION 1 AAD13895 4261595 Unknown [Saccharomyces cerevisiae] 2 AAT93224 51013861 YGL012W [Saccharomyces cerevisiae] 3 AAY42962 66270055 C24 sterol reductase [Candida glabrata] 4 XP_451223 50302575 unnamed protein product [Kluyveromyces lactis] 5 NP_986579 45201009 AGL087Cp [Eremothecium gossypii] 6 XP_456709 50407411 hypothetical protein DEHA0A08888g [Debaryomyces hansenii CBS767] 7 XP_717662 68476402 sterol C-24 (28) reductase [Candida albicans SC5314] 8 XP_503021 50551095 hypothetical protein [Yarrowia lipolytica] 9 EAU37607 114195907 Delta(24(24(1)))-sterol reductase [Aspergillus terreus NIH2624] 10 BAE61064 83770931 unnamed protein product [Aspergillus oryzae] 11 NP_594742 19115654 hypothetical protein SPAC20G4.07c [Schizosaccharomyces pombe 972h-] 12 CAA45113 5109 sts1+ [Schizosaccharomyces pombe] 13 XP_662788 67538028 hypothetical protein AN5 184.2 [Aspergillus nidulans FGSC A4] 14 XP_750461 70991224 c-24(28) sterol reductase [Aspergillus fumigatus Af293] 15 EAS37114 90307483 hypothetical protein CIMG_02468 [Coccidioides immitis RS] 16 XP_365763 39968745 hypothetical protein MG02465.4 [Magnaporthe grisea 70-15] 17 CAC18223 16944336 probable sterol C-24 reductase [Neurospora crassa] 18 XP_961419 85102913 probable sterol C-24 reductase [MIPS] [Neurospora crassa OR74A] 19 XP_660288 67524453 hypothetical protein AN2684.2 [Aspergillus nidulans FGSC A4] 20 EAU33849 114192149 Delta(24(24(1)))-sterol reductase [Aspergillus terreus NIH2624] 21 XP_753221 70996933 c-24(28) sterolreductase [Aspergillus fumigatus Af293] 22 BAE59624 83769489 unnamed protein product [Aspergillus oryzae] 23 EAT92432 111071312 predicted protein [Phaeosphaeria nodorum SN15] 24 XP_390179 46136975 hypothetical protein FG10003.1 [Gibberella zeae PH-1] 25 XP_757645 71005958 hypothetical protein UM01498.1 [Ustilago maydis 521] 26 EAS37449 90307818 hypothetical protein CIMG_02803 [Coccidioides immitis RS] 27 NP_820201 29654509 c-24(28) sterol reductase, putative [Coxiella burnetii RSA 493] 28 ZP_012 . . . 94491050 hypothetical protein CburD_01001859 [Coxiella burnetii Dugway 7E9-12] 29 AAQ88130 37039513 sterol C-24 reductase [Cryptococcus neoformans var. grubii H99] 30 XP_569697 58265082 delta24(24-1) sterol reductase [Cryptococcus neoformans var. neoformans JEC21] 31 EAQ90721 88183253 hypothetical protein CHGG_02656 [Chaetomium globosum CBS 148.51] 32 BAE55606 83765463 unnamed protein product [Aspergillus oryzae] 33 CAJ06078 68128884 sterol C-24 reductase, putative [Leishmania major] 34 ABD46555 88601114 sterol C-24 reductase-like protein [Acanthamoeba castellanii] 35 XP_638613 66809779 ERG4/ERG24 ergosterol biosynthesis protein family protein [Dictyostelium discoideum AX4]

TABLE 67 Examples of beta-carotene 15,15′-monooxygenase polypeptides. ACCESSION GI PROTEIN DESCRIPTION AAG15381 10242318 beta,beta-carotene 15,15′-dioxygenase [Mus musculus] EDL92648 149038288 beta-carotene 15,15′-monooxygenase [Rattus norvegicus] Q91XT5 46397349 Beta,beta-carotene 15,15′-monooxygenase (beta-carotene dioxygenase 1) XP_546815 57087275 PREDICTED: similar to beta,beta-carotene 15,15-monooxygenase (beta-carotene dioxygenase 1) [Canis familiaris] XP_001 . . . 149699876 PREDICTED: similar to beta,beta-carotene 15,15-dioxygenase [Equus caballus] EAW95537 119615943 beta-carotene 15,15′-monooxygenase 1, isoform CRA_a [Homo sapiens] Q9HAY6 41688803 beta,beta-carotene 15,15′-monooxygenase (beta-carotene dioxygenase 1) BAA91776 7022941 unnamed protein product [Homo sapiens] BAC35679 26344045 unnamed protein product [Mus musculus] NP_001 . . . 66792910 beta-carotene 15,15′-monooxygenase 1 [Bos taurus] XP_001 . . . 126303742 PREDICTED: similar to beta,beta-carotene 15,15-dioxygenase [Monodelphis domestica] XP_523435 114663815 PREDICTED: beta-carotene 15,15′-monooxygenase 1 [Pan troglodytes] XP_414163 50753878 PREDICTED: beta-carotene 15,15′-monooxygenase 1 [Gallus gallus] EAW95538 119615944 beta-carotene 15,15′-monooxygenase 1, isoform CRA_b [Homo sapiens] AAH56789 34784018 Zgc:63614 [Danio rerio] NP_571873 42476258 beta-carotene 15,15′-monooxygenase 1 [Danio rerio] CAC37566 13872740 putative b,b-carotene-15,15′-dioxygenase [Danio rerio] CAF95764 47214429 unnamed protein product [Tetraodon nigroviridis] CAF92469 47212035 unnamed protein product [Tetraodon nigroviridis] ABQ09267 146186430 beta-carotene 15,15′-monooxygenase 1 [Oryzias latipes] CAM14044 122890964 beta-carotene 15,15-dioxygenase 2 [Danio rerio] AAI33726 126631776 Bcdo2 protein [Rattus norvegicus] CAC37567 13872742 putative b,b-carotene-9′,10′-dioxygenase [Danio rerio] AAI07007 76825286 beta-carotene 9′,10′-dioxygenase 2 [Mus musculus] AAI35025 141795614 Bcdo2l protein [Danio rerio] NP_001 . . . 55741954 beta-carotene dioxygenase 2 [Xenopus tropicalis] XP_417929 118102019 PREDICTED: similar to carotene-9,10-monooxygenase [Gallus gallus] Q5RF16 75042584 beta,beta-carotene 9′,10′-dioxygenase (beta-carotene dioxygenase 2) (B-diox-II) XP_001 . . . 109108669 PREDICTED: beta-carotene dioxygenase 2 [Macaca mulatta] BAC41782 26449309 hypothetical protein [Macaca fascicularis] Q8HXG8 47117670 beta,beta-carotene 9′,10′-dioxygenase (beta-carotene dioxygenase 2) (B-diox-II) AAI15260 92098295 beta-carotene 15,15-dioxygenase 2 [Danio rerio] AAS20392 42560440 carotene-9′,10′-monooxygenase [Mustela putorius furo] CAG03680 47217728 unnamed protein product [Tetraodon nigroviridis] XP_536572 73955144 PREDICTED: similar to beta,beta-carotene 9,10-dioxygenase (beta- carotene dioxygenase 2) (B-diox-II) [Canis familiaris] XP_001 . . . 126327024 PREDICTED: similar to carotene-9,10-monooxygenase [Monodelphis domestica] XP_001 . . . 114640348 PREDICTED: beta-carotene dioxygenase 2 isoform 3 [Pan troglodytes] XP_001 . . . 114640346 PREDICTED: beta-carotene dioxygenase 2 isoform 4 [Pan troglodytes] AAK69433 14582265 putative carotene dioxygenase [Homo sapiens] Q9BYV7 41688802 beta,beta-carotene 9′,10′-dioxygenase (beta-carotene dioxygenase 2) (B-diox-II) EAW67194 119587598 beta-carotene dioxygenase 2, isoform CRA_f [Homo sapiens] EAW67189 119587593 beta-carotene dioxygenase 2, isoform CRA_a [Homo sapiens] XP_001 . . . 149716895 PREDICTED: similar to carotene-9,10-monooxygenase [Equus caballus] NP_001 . . . 82617628 beta-carotene dioxygenase 2 isoform b [Homo sapiens] NP_114144 82617624 beta-carotene dioxygenase 2 isoform a [Homo sapiens] CAC27994 12666531 putative b,b-carotene-9′,10′-dioxygenase [Homo sapiens] XP_508757 114640350 PREDICTED: hypothetical protein isoform 5 [Pan troglodytes] BAB55379 14042749 unnamed protein product [Homo sapiens] EAW67193 119587597 beta-carotene dioxygenase 2, isoform CRA_e [Homo sapiens] NP_001 . . . 147906096 hypothetical protein LOC791719 [Danio rerio] XP_001 . . . 125824393 PREDICTED: similar to retinal pigment epithelium 65b [Danio rerio] AAH59559 37748750 Zgc:73213 [Danio rerio] AAC14586 675485 retinal pigment epithelium-specific 61 kDa protein [Homo sapiens] XP_001 . . . 109008381 PREDICTED: retinal pigment epithelium-specific protein 65 kDa [Macaca mulatta] NP_001 . . . 71480121 retinal pigment epithelium 65b [Danio rerio] NP_084263 147902089 retinal pigment epithelium 65 [Mus musculus] AAD42042 5326773 retinal pigment epithelium-specific protein RPE65 [Cercopithecus aethiops] AAL01119 15488446 RPE65 [Mus musculus] CAA46988 564 membrane receptor p63 [Bos taurus] AAC37306 163657 retinal pigment epithelium-specific 65 kD protein EDL82582 149026339 retinal pigment epithelium 65 [Rattus norvegicus] NP_446014 16758332 retinal pigment epithelium 65 [Rattus norvegicus] NP_001 . . . 148228575 retinal pigment epithelium-specific protein 65 kDa [Xenopus laevis] AAC72356 3851555 retinal pigment epithelium-specific protein RPE65 [Canis familiaris] CAF98473 47220374 unnamed protein product [Tetraodon nigroviridis] NP_001 . . . 50978910 retinal pigment epithelium-specific protein 65 kDa [Canis lupus familiaris] AAD12758 4001821 RPE65 protein; retinal pigment epithelium 65-protein [Ambystoma tigrinum] XP_001 . . . 126305965 PREDICTED: similar to membrane receptor p63 [Monodelphis domestica] NP_001 . . . 156120623 hypothetical protein LOC514135 [Bos taurus] Q9YGX2 48475042 Retinal pigment epithelium-specific 65 kDa protein NP_001 . . . 147906384 MGC85437 protein [Xenopus laevis] NP_001 . . . 118344140 beta-carotene-15,15′-monooxygenase [Ciona intestinalis] BAC41351 26005798 retinal pigment epithelium abundant protein [Cynops pyrrhogaster] XP_001 . . . 114557125 PREDICTED: retinal pigment epithelium-specific protein 65 kDa [Pan troglodytes] NP_001 . . . 148230104 beta-carotene dioxygenase 2 [Xenopus laevis] XP_001 . . . 114640352 PREDICTED: beta-carotene dioxygenase 2 isoform 2 [Pan troglodytes] XP_001 . . . 115928922 PREDICTED: similar to beta-carotene 15,15-dioxygenase [Strongylocentrotus purpuratus] AAL39096 17432605 RPE65 [Mus musculus] XP_001 . . . 125816256 PREDICTED: similar to beta-carotene 15,15-dioxygenase 2 [Danio rerio] XP_001 . . . 156342819 hypothetical protein NEMVEDRAFT_v1g222532 [Nematostella vectensis] XP_001 . . . 125824367 PREDICTED: similar to retinal pigment epithelium 65b [Danio rerio] XP_001 . . . 156393603 predicted protein [Nematostella vectensis] XP_001 . . . 156371447 predicted protein [Nematostella vectensis] AAH67696 45767835 Bcdo2l protein [Danio rerio] CAD45010 22657452 retinal pigment epithelium-specific protein [Canis familiaris] NP_001 . . . 118344130 RPE65 homolog [Ciona intestinalis] XP_317319 118789286 ENSANGP00000006854 [Anopheles gambiae str. PEST] EAA12399 157014990 AGAP008143-PA [Anopheles gambiae str. PEST] AAY85350 68132170 carotenoid 9′,10′ monoxygenase II [Rattus norvegicus] XP_001 . . . 115969938 PREDICTED: similar to beta-carotene 15,15-dioxygenase, partial [Strongylocentrotus purpuratus] XP_001 . . . 157135394 beta-carotene dioxygenase [Aedes aegypti] XP_967460 91093114 PREDICTED: similar to CG9347-PA [Tribolium castaneum] NP_496729 71998242 Y46G5A.24 [Caenorhabditis elegans] XP_001 . . . 156398339 predicted protein [Nematostella vectensis] XP_394000 66523635 PREDICTED: similar to neither inactivation nor afterpotential B CG9347-PA isoform 1 [Apis mellifera] F88115 25395537 protein F53C3.12 [imported] - Caenorhabditis elegans AAC67462 26251528 Hypothetical protein F53C3.12 [Caenorhabditis elegans] XP_001 . . . 149709742 PREDICTED: similar to retinal pigment epithelium abundant protein RPE65 [Equus caballus] CAE71494 39582162 Hypothetical protein CBG18419 [Caenorhabditis briggsae] CAE71509 39582177 Hypothetical protein CBG18441 [Caenorhabditis briggsae] XP_001 . . . 156555525 PREDICTED: similar to beta-carotene dioxygenase [Nasonia vitripennis] XP_001 . . . 125824371 PREDICTED: similar to retinal pigment epithelium 65b [Danio rerio] CAF91639 47209442 unnamed protein product [Tetraodon nigroviridis] CAG02881 47222516 unnamed protein product [Tetraodon nigroviridis] CAG06487 47224917 unnamed protein product [Tetraodon nigroviridis] EDL95460 149041619 beta-carotene 9′,10′-dioxygenase 2 [Rattus norvegicus] YP_001 . . . 154707125 putative dioxygenase [Coxiella burnetii Dugway 7E9-12] NP_650307 24646669 neither inactivation nor afterpotential B CG9347-PA [Drosophila melanogaster] BAA94508 7593040 DRPE65 [Drosophila melanogaster] ZP_019 . . . 153206838 dioxygenase, putative [Coxiella burnetii ′MSU Goat Q177′] AAH92162 62132948 Zgc:110538 [Danio rerio] YP_444280 83814867 15,15′ beta carotene dioxygenase [Salinibacter ruber DSM 13855] BAB32248 12861637 unnamed protein product [Mus musculus] EAW67192 119587596 beta-carotene dioxygenase 2, isoform CRA_d [Homo sapiens] XP_001 . . . 114640354 PREDICTED: hypothetical protein isoform 1 [Pan troglodytes] BAC03747 21750237 unnamed protein product [Homo sapiens] EAW67190 119587594 beta-carotene dioxygenase 2, isoform CRA_b [Homo sapiens] XP_001 . . . 125774315 GA21719-PA [Drosophila pseudoobscura] YP_001 . . . 134096631 lignostilbene-alpha/ beta-dioxygenase [Saccharopolyspora erythraea NRRL 2338] EAW67191 119587595 beta-carotene dioxygenase 2, isoform CRA_c [Homo sapiens] XP_001 . . . 149486496 PREDICTED: similar to b,b-carotene-9,10-dioxygenase, partial [Ornithorhynchus anatinus] AAO90531 29541592 dioxygenase, putative [Coxiella burnetii RSA 493] XP_001 . . . 109129384 PREDICTED: similar to beta,beta-carotene 15,15-monooxygenase (beta-carotene dioxygenase 1) [Macaca mulatta] CAJ53109 109626642 beta,beta-carotene 9′,10′-dioxygenase 2 [Haloquadratum walsbyi DSM 16790] XP_001 . . . 109129778 PREDICTED: similar to beta,beta-carotene 15,15-monooxygenase (beta-carotene dioxygenase 1), partial [Macaca mulatta] XP_001 . . . 118109416 PREDICTED: similar to beta-carotene 15,15-dioxygenase, partial [Gallus gallus] ABF70124 102139989 dioxygenase-related protein [Musa balbisiana] AAW59435 57918684 decreased apical dominance protein [Petunia x hybrida] EAY74583 125526469 hypothetical protein OsI_002430 [Oryza sativa (indica cultivar-group)] AAL66961 18377622 unknown protein [Arabidopsis thaliana] EAY74584 125526470 hypothetical protein OsI_002431 [Oryza sativa (indica cultivar-group)] BAC05598 21902049 putative dioxygenase [Oryza sativa (japonica cultivar-group)] AAW33596 57116144 Dad1/CCD8 [Petunia x hybrida] NP_001 . . . 115439899 Os01g0746400 [Oryza sativa (japonica cultivar-group)] EAY75798 125527684 hypothetical protein OsI_003645 [Oryza sativa (indica cultivar-group)] EAZ13518 125572003 hypothetical protein OsJ_003343 [Oryza sativa (japonica cultivar-group)] ABQ08577 146160696 beta-carotene 15,15′-monooxygenase 2 [Oryzias latipes] BAB63485 15289786 retinal pigment epithelium 65-like [Oryza sativa (japonica cultivar-group)] AAS66906 45504725 dioxygenase RAMOSUS1 [Pisum sativum] ZP_013 . . . 95927318 hypothetical protein CburR_01000925 [Coxiella burnetii RSA 331] YP_658115 110668304 beta,beta-carotene 15,15′-monooxygenase; beta-carotene dioxygenase 1 [Haloquatratum walsbyi DSM 16790] CAL90971 121495644 torulene oxygenase [Gibberella fujikuroi] YP_134677 55376826 retinal pigment epithelial membrane protein [Haloarcula marismortui ATCC 43049] CAL90970 121495642 torulene oxygenase [Gibberella fujikuroi] XP_382801 46111487 hypothetical protein FG02625.1 [Gibberella zeae PH-1] XP_001 . . . 149480289 PREDICTED: similar to carotene-9,10-monooxygenase, partial [Ornithorhynchus anatinus] ABQ76053 148508268 beta,beta-carotene 9′,10′-dioxygenase 2 [uncultured haloarchaeon] YP_657779 110667968 beta,beta-carotene 9′,10′-dioxygenase 2 [Haloquatratum walsbyi DSM 16790] XP_001 . . . 109149184 PREDICTED: similar to beta,beta-carotene 15,15-monooxygenase (beta-carotene dioxygenase 1), partial [Macaca mulatta] AAP92577 33086530 Ab2-079 [Rattus norvegicus] ABN50352 125662819 carotenoid cleavage dioxygenase 1 [Hypcrea jecorina] EAL93736 66853412 dioxygenase, putative [Aspergillus fumigatus Af293] XP_001 . . . 125876361 PREDICTED: hypothetical protein, partial [Danio rerio] XP_001 . . . 119481741 dioxygenase, putative [Neosartorya fischeri NRRL 181] XP_001 . . . 119494253 retinal pigment epithelial membrane family protein [Neosartorya fischeri NRRL 181] XP_958452 85090512 hypothetical protein [Neurospora crassa OR74A] ED064867 157069539 predicted protein [Neurospora crassa OR74A] XP_001 . . . 121716178 dioxygenase, putative [Aspergillus clavatus NRRL 1] CAL64769 148353839 putative carotene oxygenase [Blakeslea trispora] BAD79505 56686283 lignostilbene-alpha beta-dioxygenase [Synechococcus elongatus PCC 6301] ZP_016 . . . 119509407 Retinal pigment epithelial membrane protein [Nodularia spumigena CCY9414] BAE56886 83766746 unnamed protein product [Aspergillus oryzae] BAE59247 83769110 unnamed protein product [Aspergillus oryzae] ECC98172 139038333 hypothetical protein GOS_5249294 [marine metagenome] ZP_017 . . . 126656631 beta-carotene 15,15′-dioxygenase [Cyanothece sp. CCY0110] BAC91630 35214261 gll3689 [Gloeobacter violaceus PCC 7421] EAT90277 111069157 hypothetical protein SNOG_02065 [Phaeosphaeria nodorum SN15] CAB79998 7270228 putative protein [Arabidopsis thaliana] CAL49287 117557505 putative carotene-dioxygenase [Rhizopus oryzae] XP_659308 67522495 hypothetical protein AN1704.2 [Aspergillus nidulans FGSC A4] XP_001 . . . 125824373 PREDICTED: similar to retinal pigment epithelium 65b [Danio rerio] XP_001 . . . 119185939 hypothetical protein CIMG_03017 [Coccidioides immitis RS] 2BIWB 66361521 Chain B, Crystal Structure Of Apocarotenoid Cleavage Oxygenase From Synechocystis, Native Enzyme ABA20860 75701184 Retinal pigment epithelial membrane protein [Anabaena variabilis ATCC 29413] XP_001 . . . 116207620 hypothetical protein CHGG_03103 [Chaetomium globosum CBS 148.51] XP_001 . . . 145257182 hypothetical protein An04g02710 [Aspergillus niger] BAB75983 17133419 lignostilbene-alpha, beta-dioxygenase [Nostoc sp. PCC 7120] XP_501958 50548975 hypothetical protein [Yarrowia lipolytica] XP_001 . . . 109129782 PREDICTED: similar to beta,beta-carotene 15,15-monooxygenase (beta-carotene dioxygenase 1), partial [Macaca mulatta] ZP_016 . . . 119484919 Retinal pigment epithelial membrane protein [Lyngbya sp. PCC 8106] ZP_017 . . . 126656882 Retinal pigment epithelial membrane protein [Cyanothece sp. CCY0110] XP_001 . . . 109149035 PREDICTED: similar to beta,beta-carotene 15,15-monooxygenase (beta-carotene dioxygenase 1), partial [Macaca mulatta] CAL49288 117557507 putative carotene-dioxygenase [Rhizopus oryzae] ZP_001 . . . 23130610 COG3670: Lignostilbene-alpha, beta-dioxygenase and related enzymes [Nostoc punctiforme PCC 73102] XP_362433 39945792 hypothetical protein MGG_08016 [Magnaporthe grisea 70-15] XP_001 . . . 115951205 PREDICTED: hypothetical protein [Strongylocentrotus purpuratus] AAC12875 3033545 lignostilbene-alpha, beta-dioxygenase [Synechococcus sp. PCC 7942] XP_001 . . . 154282379 conserved hypothetical protein [Ajellomyces capsulatus NAm1] ABG52331 110167791 Carotenoid oxygenase[Trichodesmium erythraeum IM S101] ABC99263 86554305 putative lignostilbene-alpha, beta-dioxygenase [Synechococcus sp. JA-3-3Ab] NP_931511 37528166 hypothetical protein plu 4336 [Photorhabdus luminescens subsp. laumondii TTO1] EDD33453 143171893 hypothetical protein GOS_1320815 [marine metagenome] XP_001 . . . 115387251 predicted protein [Aspergillus terreus NIH2624] YP_885943 118467371 lignostilbene-alpha, beta-dioxygenase [Mycobacterium smegmatis str. MC2 155]

TABLE 68 Examples of beta-carotene retinol dehydrogenase polypeptides. ACCESSION GI PROTEIN DESCRIPTION AAX33670 60678607 epithelial retinol dehydrogenase [Mus musculus] BAC34110 26340896 unnamed protein product [Mus musculus] XP_001 . . . 149730699 PREDICTED: similar to short-chain dehydrogenase/reductase RETSDR8 [Equus caballus] XP_545513 74004823 PREDICTED: similar to NADP-dependent retinol dehydrogenase/reductase [Canis familiaris] Q8HYR6 75064969 Dehydrogenase/reductase SDR family member 9 precursor (3-alpha hydroxysteroid dehydrogenase) (3 alpha-HSD) (Short-chain dehydrogenase/reductase retSDR8) XP_001 . . . 114581559 PREDICTED: NADP-dependent retinol dehydrogenase/reductase isoform 3 [Pan troglodytes] XP_001 . . . 109099963 PREDICTED: similar to NADP-dependent retinol dehydrogenase/reductase isoform 3 [Macaca mulatta] AAL37037 17224600 short-chain dehydrogenase/reductase RETSDR8 [Homo sapiens] AAD32458 4894382 retinol dehydrogenase homolog [Homo sapiens] XP_001 . . . 126326289 PREDICTED: similar to short-chain dehydrogenase/reductase RETSDR8 [Monodelphis domestica] XP_001 . . . 149639643 PREDICTED: similar to short-chain dehydrogenase/reductase RETSDR8 [Ornithorhynchus anatinus] XP_001 . . . 114581568 PREDICTED: NADP-dependent retinol dehydrogenase/reductase isoform 1 [Pan troglodytes] AAF82748 9082139 retinol dehydrogenase homolog isoform-1 [Homo sapiens] XP_001 . . . 109099969 PREDICTED: similar to NADP-dependent retinol dehydrogenase/reductase isoform 1 [Macaca mulatta] AAY24306 62988919 unknown [Homo sapiens] XP_422015 118093666 PREDICTED: similar to short-chain dehydrogenase/reductase RETSDR8 [Gallus gallus] XP_001 . . . 114581566 PREDICTED: NADP-dependent retinol dehydrogenase/reductase isoform 2 [Pan troglodytes] NP_001 . . . 147905288 NADP-dependent retinol dehydrogenase/reductase [Xenopus laevis] AAH67304 45501367 Dehydrogenase/reductase (SDR family) member 9 [Xenopus tropicalis] EDM16447 149066574 rCG60176, isoform CRA_a [Rattus norvegicus] AAH62000 38303814 Rdh2 protein [Rattus norvegicus] AAC52316 1072046 retinol dehydrogenase type II NP_001 . . . 148227174 hypothetical protein LOC398882 [Xenopus laevis] NP_033066 6677697 retinol dehydrogenase 16 [Mus musculus] NP_001 . . . 147900576 hypothetical protein LOC432178 [Xenopus laevis] NP_694773 23680945 retinol dehydrogenase 9 [Mus musculus] NP_003716 19743808 hydroxysteroid (17-beta) dehydrogenase 6 [Homo sapiens] BAE21017 74205129 unnamed protein product [Mus musculus] EDL24523 148692576 mCG140848 [Mus musculus] AAL14860 22651436 cis-retinol/3 alpha hydroxysterol short-chain dehydrogenase-like protein [Mus musculus] P55006 1710631 Retinol dehydrogenase 7 (Retinol dehydrogenase type III) (RODH III) AAH82896 52354822 LOC494781 protein [Xenopus laevis] XP_001 . . . 109097334 PREDICTED: similar to 3-hydroxysteroid epimerase [Macaca mulatta] AAI35515 134023801 LOC100124842 protein [Xenopus tropicalis] P50169 1710629 Retinol dehydrogenase 3 (Retinol dehydrogenase type I) (RODH I) BAC98304 37360649 cis-retinol/androgen dehydrogenase type 3 [Mus musculus] NP_663399 21703846 cis-retinol/3 alpha hydroxysterol short-chain dehydrogenase-like [Mus musculus] AAB07997 841197 retinol dehydrogenase type I XP_001 . . . 126343946 PREDICTED: similar to microsomal NAD+-dependent retinol dehydrogenase 4 [Monodelphis domestica] NP_775427 27545384 hydroxysteroid (17-beta) dehydrogenase 6 [Rattus norvegicus] EDL24527 148692580 mCG134493 [Mus musculus] XP_531641 73968454 PREDICTED: similar to microsomal NAD+-dependent retinol dehydrogenase 4 isoform 1 [Canis familiaris] AAH24603 19354049 Retinol dehydrogenase 7 [Mus musculus] AAL02134 22074192 retinol dehydrogenase similar protein 2 [Mus musculus] NP_536684 20147789 retinol dehydrogenase 1 (all trans) [Mus musculus] AAH88104 56789874 Hydroxysteroid (17-beta) dehydrogenase 6 [Rattus norvegicus] EDL24532 148692585 hydroxysteroid (17-beta) dehydrogenase 9, isoform CRA_a [Mus musculus] ABG81445 110665598 microsomal NAD+-dependent retinol dehydrogenase 4 [Bos taurus] EDL24533 148692586 hydroxysteroid (17-beta) dehydrogenase 9, isoform CRA_b [Mus musculus] XP_538239 73968448 PREDICTED: similar to 3-hydroxysteroid epimerase isoform 3 [Canis familiaris] CAJ81333 89272084 novel retinol dehydrogenase protein [Xenopus tropicalis] AAH46694 28302191 Rodh-A-prov protein [Xenopus laevis] XP_001 . . . 149715148 PREDICTED: similar to microsomal NAD+-dependent retinol dehydrogenase 4 [Equus caballus] XP_001 . . . 109097338 PREDICTED: similar to microsomal NAD+-dependent retinol dehydrogenase 4 isoform 2 [Macaca mulatta] XP_001 . . . 109097336 PREDICTED: similar to microsomal NAD+-dependent retinol dehydrogenase 4 isoform 3 [Macaca mulatta] XP_858182 73968456 PREDICTED: similar to microsomal NAD+-dependent retinol dehydrogenase 4 isoform 2 [Canis familiaris] AAI02627 74354657 Similar to 3-hydroxysteroid epimerase [Bos taurus] XP_858059 73968446 PREDICTED: similar to 3-hydroxysteroid epimerase isoform 4 [Canis familiaris] AAC39922 3372592 sterol/retinol dehydrogenase [Homo sapiens] AAY44823 66354281 retinol dehydrogenase 5 [Danio rerio] XP_522604 55639443 PREDICTED: retinol dehydrogenase 16 [Pan troglodytes] AAH97151 66911361 Retinol dehydrogenase 5 (11-cis/9-cis) [Danio rerio] CAK05049 94732559 novel protein similar to vertebrate retinol dehydrogenase family [Danio rerio] NP_990044 45382649 11-cis retinol dehydrogenase [Gallus gallus] EAW96968 119617374 retinol dehydrogenase 16 (all-trans and 13-cis) [Homo sapiens] NP_001 . . . 148233310 hypothetical protein LOC398939 [Xenopus laevis] CAG04959 47218630 unnamed protein product [Tetraodon nigroviridis] AAH88014 56789076 Hypothetical LOC496749 [Xenopus tropicalis] AAI23192 114107846 Rdh9 protein [Xenopus laevis] AAC72923 3859946 retinol dehydrogenase [Homo sapiens] AAF82747 9082137 retinol dehydrogenase homolog isoform-2 [Homo sapiens] AAH75455 49522586 MGC89248 protein [Xenopus tropicalis] AAH59614 37590384 Retinol dehydrogenase 1, like [Danio rerio] AAB88252 2661211 oxidative 3 alpha hydroxysteroid dehydrogenase [Homo sapiens] EDL24633 148692686 retinol dehydrogenase 5, isoform CRA_b [Mus musculus] BAE22435 74149341 unnamed protein product [Mus musculus] XP_538220 73968269 PREDICTED: similar to 11-cis retinol dehydrogenase (11-cis RDH) [Canis familiaris] NP_001 . . . 147899633 MGC84134 protein [Xenopus laevis] XP_001 . . . 109097121 PREDICTED: retinol dehydrogenase 5 (11-cis and 9-cis) [Macaca mulatta] NP_001 . . . 147905378 MGC84099 protein [Xenopus laevis] CAF92451 47210426 unnamed protein product [Tetraodon nigroviridis] XP_522429 55638373 PREDICTED: similar to 11-cis retinol dehydrogenase isoform 2 [Pan troglodytes] AAH88529 56788840 Hypothetical LOC496830 [Xenopus tropicalis] NP_002896 50726952 retinol dehydrogenase 5 (11-cis and 9-cis) [Homo sapiens] CAA57715 663171 11-cis retinol dehydrogenase [Bos taurus] AAH28298 20271410 Retinol dehydrogenase 5 (11-cis/9-cis) [Homo sapiens] NP_001 . . . 148230094 hypothetical protein LOC399015 [Xenopus laevis] AAA80694 1054531 11-cis-retinol dehydrogenase NP_932335 37620196 retinol dehydrogenase 1 [Danio rerio] XP_538240 73968424 PREDICTED: similar to orphan short-chain dehydrogenase/ reductase [Canis familiaris] AAI34659 134024746 MGC151815 protein [Bos taurus] AAH64820 40673988 Orphan short chain dehydrogenase/reductase [Mus musculus] EDL24531 148692584 orphan short chain dehydrogenase/reductase [Mus musculus] AAB93668 1916935 9-cis-retinol specific dehydrogenase [Homo sapiens] XP_001 . . . 149715290 PREDICTED: hypothetical protein [Equus caballus] XP_001 . . . 109482100 PREDICTED: similar to retinol dehydrogenase similar protein [Rattus norvegicus] EAW96967 119617373 orphan short-chain dehydrogenase/reductase [Homo sapiens] XP_522439 114644207 PREDICTED: orphan short-chain dehydrogenase/reductase [Pan troglodytes] EDL24528 148692581 mCG134494, isoform CRA_a [Mus musculus] ABQ09278 146186452 retinol dehydrogenase 1 [Oryzias latipes] ABQ09279 146186454 retinol dehydrogenase 5 [Oryzias latipes] XP_001 . . . 109097340 PREDICTED: similar to microsomal NAD+-dependent retinol dehydrogenase 4 isoform 1 [Macaca mulatta] EDM16444 149066571 orphan short chain dehydrogenase/reductase [Rattus norvegicus] AAG44849 12006418 microsomal retinol dehydrogenase [Branchiostoma lanceolatum] BAE88760 90078160 unnamed protein product [Macaca fascicularis] AAG44847 12006414 microsomal retinol dehydrogenase 1 [Branchiostoma floridae] XP_782726 115665382 PREDICTED: similar to retinol dehydrogenase type 1, partial [Strongylocentrotus purpuratus] XP_001 . . . 115939186 PREDICTED: similar to retinol dehydrogenase type 1 [Strongylocentrotus purpuratus] XP_001 . . . 118093664 PREDICTED: similar to MGC83505 protein [Gallus gallus] XP_781379 115654644 PREDICTED: similar to retinol dehydrogenase type 1, partial [Strongylocentrotus purpuratus] XP_001 . . . 115958895 PREDICTED: similar to retinol dehydrogenase type 1 [Strongylocentrotus purpuratus] AAG44848 12006416 microsomal retinol dehydrogenase 2 [Branchiostoma floridae] XP_001 . . . 115960451 PREDICTED: similar to retinol dehydrogenase type III [Strongylocentrotus purpuratus] EDL84786 149029615 rCG42422, isoform CRA_a [Rattus norvegicus] AAL14861 22651438 cis-retinol/3 alpha hydroxysterol short-chain dehydrogenase-like protein isoform [Mus musculus] XP_001 . . . 156386264 predicted protein [Nematostella vectensis] XP_001 . . . 156389613 predicted protein [Nematostella vectensis] XP_001 . . . 156368581 predicted protein [Nematostella vectensis] XP_001 . . . 156362183 predicted protein [Nematostella vectensis] BAC28142 26328807 unnamed protein product [Mus musculus] XP_001 . . . 156367012 predicted protein [Nematostella vectensis] XP_001 . . . 156367010 predicted protein [Nematostella vectensis] CAG14527 47203967 unnamed protein product [Tetraodon nigroviridis] XP_001 . . . 156367016 predicted protein [Nematostella vectensis] XP_001 . . . 156401057 predicted protein [Nematostella vectensis] AAB69884 2384782 Dehydrogenases, short chain protein 16 [Caenorhabditis elegans] XP_001 . . . 156367014 predicted protein [Nematostella vectensis] XP_001 . . . 156367008 predicted protein [Nematostella vectensis] NP_871815 32563809 DeHydrogenases, Short chain family member (dhs-2) [Caenorhabditis elegans] NP_491575 17507613 DeHydrogenases, Short chain family member (dhs-2) [Caenorhabditis elegans] NP_505941 115534660 DeHydrogenases, Short chain family member (dhs-20) [Caenorhabditis elegans] CAE74886 39583813 Hypothetical protein CBG22749 [Caenorhabditis briggsae] XP_001 . . . 156408325 predicted protein [Nematostella vectensis] XP_001 . . . 126343453 PREDICTED: similar to 3-hydroxybutyrate dehydrogenase, type 1 [Monodelphis domestica] CAE64533 39593064 Hypothetical protein CBG09275 [Caenorhabditis briggsae] XP_001 . . . 149571441 PREDICTED: similar to retinol dehydrogenase similar protein, partial [Ornithorhynchus anatinus] NP_955017 49227085 truncated cis-retinol/3 alpha-hydroxysterol short-chain dehydrogenase [Mus musculus] XP_001 . . . 156363786 predicted protein [Nematostella vectensis] XP_001 . . . 109097342 PREDICTED: similar to microsomal NAD+-dependent retinol dehydrogenase 4 [Macaca mulatta] NP_001 . . . 147899736 hypothetical protein LOC100037356 [Danio rerio] XP_001 . . . 156366980 predicted protein [Nematostella vectensis] XP_001 . . . 125853068 PREDICTED: hypothetical protein [Danio rerio] AAH11964 15080429 3-hydroxybutyrate dehydrogenase, type 1 [Homo sapiens] XP_345771 109480152 PREDICTED: similar to retinol dehydrogenase 5 [Rattus norvegicus] XP_001 . . . 149731507 PREDICTED: similar to 3-hydroxybutyrate dehydrogenase, type 1 [Equus caballus] NP_976059 44680136 3-hydroxybutyrate dehydrogenase precursor [Homo sapiens] XP_001 . . . 149633623 PREDICTED: similar to 3-hydroxybutyrate dehydrogenase, type 1 [Ornithorhynchus anatinus] AAH89597 58476255 BC089597 protein [Mus musculus] XP_001 . . . 114591350 PREDICTED: 3-hydroxybutyrate dehydrogenase isoform 1 [Pan troglodytes] XP_516981 114591348 PREDICTED: 3-hydroxybutyrate dehydrogenase isoform 2 [Pan troglodytes] XP_001 . . . 156405501 predicted protein [Nematostella vectensis] CAG04267 47223406 unnamed protein product [Tetraodon nigroviridis] XP_545160 74002972 PREDICTED: similar to 3-hydroxybutyrate dehydrogenase precursor [Canis familiaris] AAI03052 73587365 3-hydroxybutyrate dehydrogenase, type 1 [Bos taurus] BAF35260 116267595 11 beta-hydroxysteroid dehydrogenase short form [Anguilla japonica] AAH65613 41350974 Hydroxysteroid 11-beta dehydrogenase 2 [Danio rerio] AAB59684 203921 D-beta-hydroxybutyrate dehydrogenase BAE27076 74226863 unnamed protein product [Mus musculus] NP_446447 55742813 3-hydroxybutyrate dehydrogenase, type 1 [Rattus norvegicus] P29147 68837285 D-beta-hydroxybutyrate dehydrogenase, mitochondrial precursor (BDH) (3-hydroxybutyrate dehydrogenase) EDK97750 148665334 3-hydroxybutyrate dehydrogenase, type 1 [Mus musculus] CAE75331 39592111 Hypothetical protein CBG23309 [Caenorhabditis briggsae] BAE23523 74206030 unnamed protein product [Mus musculus] BAC36453 26345604 unnamed protein product [Mus musculus] ABH02937 110931880 17-beta hydroxysteroid dehydrogenase 2 [Bos taurus] NP_509415 17568189 F55E10.6 [Caenorhabditis elegans] XP_546810 73957269 PREDICTED: similar to hydroxysteroid (17-beta) dehydrogenase 2 [Canis familiaris] XP_001 . . . 115925077 PREDICTED: similar to retinol dehydrogenase type 1 [Strongylocentrotus purpuratus] AAA58352 177198 (R)-3-hydroxybutyrate dehydrogenase XP_422703 50752249 PREDICTED: hypothetical protein [Gallus gallus] XP_001 . . . 149640222 PREDICTED: similar to 17-beta hydroxysteroid dehydrogenase 2 [Ornithorhynchus anatinus] XP_511130 114663836 PREDICTED: hydroxysteroid (17-beta) dehydrogenase 2 [Pan troglodytes] P37059 544152 Estradiol 17-beta-dehydrogenase 2 (17-beta-HSD 2) (Microsomal 17-beta-hydroxysteroid dehydrogenase) (20 alpha-hydroxysteroid dehydrogenase) (20-alpha-HSD) (E2DH) AAV38556 54696368 hydroxysteroid (17-beta) dehydrogenase 2 [synthetic construct] AAH90448 60649586 Hsd17b2 protein [Danio rerio] XP_001 . . . 118087716 PREDICTED: hypothetical protein [Gallus gallus] NP_001 . . . 93277072 hydroxysteroid (17-beta) dehydrogenase 2[Danio rerio] P51658 2507561 Estradiol 17-beta-dehydrogenase 2 (17-beta-HSD 2) (17-beta- hydroxysteroid dehydrogenase 2) BAC76709 30844232 11-beta-hydroxysteroid dehydrogenase [Oncorhynchus mykiss] AAY84567 67975199 17-beta hydroxysteroid dehydrogenase 2 [Macaca fascicularis] BAD96721 62897563 hydroxysteroid (17-beta) dehydrogenase 2 variant [Homo sapiens] XP_414168 118096516 PREDICTED: similar to 17 beta hydroxysteroid dehydrogenase type 2 [Gallus gallus] CAE66933 39589698 Hypothetical protein CBG12325 [Caenorhabditis briggsae] AAK95857 22074155 retinol dehydrogenase similar protein [Rattus norvegicus] AAH74144 49257832 MGC81883 protein [Xenopus laevis] XP_001 . . . 125850087 PREDICTED: hypothetical protein [Danio rerio] AAH90181 58701977 Zgc:113142 [Danio rerio] AAA87007 755574 11-beta-hydroxylsteroid dehydrogenase type 2 Q62730 3334165 Estradiol 17-beta-dehydrogenase 2 (17-beta-HSD 2) (17-beta- hydroxysteroid dehydrogenase 2) XP_001 . . . 115956172 PREDICTED: similar to Retinol dehydrogenase 3 (Retinol dehydrogenase type I) (RODH I) [Strongylocentrotus purpuratus] CAA64982 1200097 17-beta-hydroxysteroid dehydrogenase type II [Mus musculus] EDL11587 148679640 hydroxysteroid (17-beta) dehydrogenase 2 [Mus musculus] EDL92656 149038296 hydroxysteroid (17-beta) dehydrogenase 2 [Rattus norvegicus] XP_001 . . . 149626583 PREDICTED: similar to 11 beta-hydroxysteroid dehydrogenase type 2 [Ornithorhynchus anatinus] XP_001 . . . 109482098 PREDICTED: similar to Retinol dehydrogenase 2 (Retinol dehydrogenase type II) (RODH II) (29 k-protein) [Rattus norvegicus] XP_001 . . . 109128927 PREDICTED: similar to Corticosteroid 11-beta-dehydrogenase isozyme 2 (11-DH2) (11-beta-hydroxysteroid dehydrogenase type 2) (11-beta-HSD2) (NAD-dependent 11-beta-hydroxysteroid dehydrogenase) [Macaca mulatta]

TABLE 69 Genes Comprising the SAGA Complex. S. cerevisiae gene Y lipolytica gene SPT3 YALI0E21417g SPT7 none identified SPT8 YALI0E23804g SPT20 YALI0C04070g ADA2 YALI0F14443g ADA3 none identified GCN5 YALI0E02772g ADA1 none identified TRA1 YALI0C02057g

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. The scope of the present invention is not intended to be limited to the above Description, but rather is as set forth in the following claims: 

The invention claimed is:
 1. An engineered Saccharomyces cerevisiae strain that produces at least one retinolic compound selected from the group consisting of retinol, retinal, retinoic acid, retinyl palmitate and combinations thereof, the strain comprising at least one retinologenic modification selected from the group consisting of increased expression or activity of a beta-carotene 15,15′-monooxygenase polypeptide, increased expression or activity of a retinol dehydrogenase polypeptide, and combinations thereof; wherein the engineered strain can accumulate lipid to at least about 20% of its dry cell weight; and wherein as a result of genetic engineering, the engineered strain produces the at least one retinolic compound to a level at least about 1% of its dry cell weight.
 2. The engineered Saccharomyces cerevisiae strain of claim 1, wherein the strain does not naturally produce the at least one retinolic compound.
 3. The engineered Saccharomyces cerevisiae strain of claim 1, further comprising at least one oleaginic modification.
 4. The engineered Saccharomyces cerevisiae strain of claim 3, wherein the at least one oleaginic modification increases or decreases expression or activity of at least one oleaginic polypeptide, selected from the group consisting of acetyl-CoA carboxylase polypeptide, pyruvate decarboxylase polypeptide, isocitrate dehydrogenase polypeptide, ATP-citrate lyase polypeptide, malic enzyme polypeptide, AMP deaminase polypeptide, malate dehydrogenase polypeptide, glucose-6-phosphate dehydrogenase polypeptide, 6-phosphogluconate dehydrogenase polypeptide, fructose 1,6 bisphosphatase polypeptide, NADH kinase polypeptide, transhydrogenase polypeptide, acyl-CoA:diacylglycerol acyltransferase polypeptide, phospholipid:diacylglycerol acyltransferase polypeptide, acyl-CoA:cholesterol acyltransferase polypeptide, triglyceride lipase polypeptide, acyl-coenzyme A oxidase polypeptide and combinations thereof.
 5. The engineered Saccharomyces cerevisiae strain of claim 3, wherein the at least one oleaginic modification increases or decreases expression or activity of at least one oleaginic polypeptide selected from the group consisting of a polypeptide in any one of Tables 1 through 6 or 31-47.
 6. The engineered Saccharomyces cerevisiae strain of claim 1, wherein the at least one retinologenic modification confers to the engineered strain the ability to produce the at least one retinolic compound to a level at least about 1% of its dry cell weight.
 7. The engineered Saccharomyces cerevisiae strain of claim 1, wherein the at least one retinologenic modification confers to the engineered strain the ability to produce at least one retinolic compound which a non-engineered strain does not naturally produce.
 8. The engineered Saccharomyces cerevisiae strain of claim 1, wherein the at least one retinologenic modification increases expression or activity of beta-carotene 15,15′-monoxygenase.
 9. The engineered Saccharomyces cerevisiae strain of claim 1, wherein the beta-carotene 15,15′-monooxygenase polypeptide is selected from the group consisting of a polypeptide in Table
 67. 10. The engineered Saccharomyces cerevisiae strain of claim 1, wherein the at least one retinologenic modification further comprises decreasing the expression or activity of one or more components of the SAGA histone acetyltransferase complex.
 11. The engineered Saccharomyces cerevisiae strain of claim 10, wherein the at least one retinologenic modification comprises disruption of the endogenous SPT8 gene.
 12. The engineered Saccharomyces cerevisiae strain of claim 10, wherein the one or more components of the SAGA histone acetyltransferase complex are selected from the polypeptides listed in Table
 69. 13. The engineered Saccharomyces cerevisiae strain of claim 1, wherein the retinologenic modification further comprises altering the expression or activity of one or more carotenoid biosynthesis polypeptides.
 14. The engineered Saccharomyces cerevisiae strain of claim 13, wherein the one or more carotenoid biosynthesis polypeptides are selected from the group consisting of phytoene synthase, phytoene dehydrogenase, lycopene cyclase, carotene ketolase, carotene hydroxylase, astaxanthin synthase, carotenoid epsilon hydroxylase, lycopene cyclase, carotenoid glucosyltransferase, acyl CoA:diacyglycerol acyltransferase, geranylgeranylpyrophosphate synthase, and truncated HMG-CoA reductase.
 15. The engineered Saccharomyces cerevisiae strain of claim 13, wherein the one or more carotenoid biosynthesis polypeptides are selected from the sequences provided in Tables 17a-25.
 16. A method of producing a retinolic compound, the method comprising steps of: a. cultivating the fungus of claim 1 under conditions that allow production of the retinolic compound; b. and isolating the produced retinolic compound.
 17. The engineered Saccharomyces cerevisiae strain of claim 1, wherein the at least one retinologenic modification comprises introduction of a gene encoding a heterologous beta-carotene 15,15′-monooxygenase polypeptide, a heterologous retinol dehydrogenase polypeptide, or combinations thereof.
 18. The engineered Saccharomyces cerevisiae strain of claim 1, wherein the retinol dehydrogenase polypeptide is selected from the group consisting of a polypeptide in Table
 68. 19. An engineered Saccharomyces cerevisiae strain that produces at least one retinolic compound selected from the group consisting of retinol, retinal, retinoic acid, retinyl palmitate and combinations thereof, the strain comprising at least one genetic modification selected from the group consisting of: increased expression or activity of a beta-carotene 15,15′-monooxygenase polypeptide; increased expression or activity of a retinol dehydrogenase polypeptide; increased expression or activity of phytoene synthase; increased expression or activity of lycopene cyclase; increased expression or activity of phytoene dehydrogenase; increased expression or activity of geranylgeranylpyrophosphate synthase; increased expression or activity of truncated HMG-CoA reductase; and combinations thereof; wherein the engineered strain can accumulate lipid to at least about 20% of its dry cell weight; and wherein as a result of genetic engineering, the engineered strain produces the at least one retinolic compound to a level at least about 1% of its dry cell weight.
 20. The engineered Saccharomyces cerevisiae strain of claim 19, wherein the genetic modification comprises introduction of one or more heterologous genes encoding metabolic enzymes selected from the group consisting of: beta-carotene 15,15′-monooxygenase, phytoene synthase, lycopene cyclase, phytoene dehydrogenase, geranylgeranylpyrophosphate synthase, truncated HMG-CoA reductase, and combinations thereof. 